이 작품은 과학진흥진흥협회(JSPS) 과학연구부(C) 18K08956과 후쿠오카대학 중앙연구원(201045)의 기금을 지원받았다.

이 연구는 일본 후쿠오카대학(IRB No. 2017M017)의 기관심사위원회(IRB)의 승인을 받았습니다. 참여 하기 전에, 모든 환자는 서면 통보 된 동의를 제공.
1. fNIRS 실험 준비
참고: 이 실험을 위한 다중 채널 연속 파 레이저 기반 NIRS 시스템이 사용되었습니다. 근적외선의 파장은 780nm, 805nm, 830nm였으며 샘플링 속도는 7.8Hz로 설정되었다. 시간 및 공간 해상도(광 방출기와 검출기 프로브 사이의 거리)는 각각 0.13 s및 3.0cm였습니다.
<ol><li>fNIRS 장치를 어두운 소음이 없는 장소에 설정합니다. 실온에서 실험을 수행합니다. 실험 30분 전에 fNIRS 계측기를 시작합니다. 참고: fNIRS 시스템은 제어된 온도 및 습도 조건에서 사용되어장비(26)의안정적인 작동을 보장합니다.</li>
	<li>fNIRS 레코딩을 위해 전체 헤드 캡을 사용하고 국제 10-20 시스템의 중앙(Cz)에 해당하는 위치가 헤드 캡의 홀더 245에 위치하도록 피사체의 머리에 헤드 캡을 부착한다. (그림1).</li>
	<li>마킹 스티커를 참조 위치 지점에 부착합니다: nasion (Nz), 오른쪽 외부 청각 육류 (AR), 왼쪽 외부 청각 고기 (AL). 참고: 3차원(3D) 좌표는 Nz, AR, AL 마킹 스티커 및 Cz 홀더의 위치를 중심으로 판독되므로 사진을 찍기 전에 표시 스티커를 부착해야 합니다.</li>
	<li>공간 등록을 위한 고해상도 디지털 카메라 보정 후 15개의 관점에서 참조점(Cz, Nz, AR 및 LR)을 표시하면서 프로브 위치로 피사체의 머리를 촬영합니다. 참고: 프로브를 배치하기 전에 사진을 찍어주세요. 프로브를 배치한 후 사진을 찍은 경우 마커 랜드마크가 프로브 및 배선 코드에 의해 숨겨질 수 있습니다. 제조업체가 권장하는 대로 피사체 오른쪽으로 대각선으로 30° 12장의 사진을 찍은 후, 3장 이상의 사진을 사진 위에 약간 가져가서 Cz(홀더 245호)이 그림에 나타납니다. 총 15회 이상의 샷을 캡처할 때 입체화가 용이하기 때문입니다.</li>
	<li>발광 다이오드(LED)-조명 플라스틱 막대를 사용하여 검안장치를 방해하는 피사체의 머리카락을 조심스럽게 분리하여 프로브를 부착합니다. 검소가 두피 표면에서 최소한의 거리와 두피와 접촉하도록 프로브를 정렬합니다. 참고: 자율신경계활성화(23)와관련된 전신 confounders의 강도 증가로 인해 검안의 부착으로 인해 환자에게 압력이나 불편함이 있는지 신중하게 확인한다.</li>
	<li>48채널 시스템을 32개의 광원(광원 16개, 검출기 16개, 각 반구당 4x4어레이)을 관심 영역으로 정면 및 정수리 영역을 양측으로 헤드 캡에 배치한다(그림2).</li>
	<li>3D 디지타이저 소프트웨어를 시작하고 사용하여 공간 등록을 결정합니다.</li>
	<li>전체 헤드의 그림 데이터를 스캔한 후 Origin 및 기타 파일(*CSV 파일)으로 자동 측정하여 각 환자의 공간 좌표를 결정합니다. 참고: 자동 측정을 사용하여 이미지에서 좌표 점을 감지할 수 없는 경우 수동으로 조정을 입력합니다.</li>
</ol>2. 실험 실행
<ol><li>실험을 위한 블록 설계를 선택하고, 손 개폐, 손가락 태핑 등과 같은 연구에 대한 관심의 움직임일 수 있다. 우리의 이전 연구에서, 작업은 로봇 보조 팔꿈치운동이었다 15. 참고: 각 주기는 3개의 블록(15s의 나머지 - 15s의 작업 - 15s 의 나머지)으로 만들어지며 각 환자는 각 세션에서 7 개의 주기를 완료합니다.</li>
	<li>참가자가 시작 신호가 될 때까지 편안한 위치에서 기다립니다. 나머지와 작업 중에 피사체에게 눈을 감도록 지시한다.</li>
	<li>시작 및 중지 큐를 제공합니다 (즉, "팔꿈치의 반복 굴곡과 확장", "중지하고 휴식"). 참고: 측정 중에 말하지 마십시오. 측정 중에 모니터 화면에서 아티팩트를 주의 깊게 확인합니다.</li>
	<li>블록 설계 작업을 동일한 자세로 수행합니다. 서 있거나 앉는 자세를 가진 직립 자세는 헤드셋을 왜곡하지 않는 것이 바람직합니다. 참고: 환자가 장시간 머리 홀더를 착용한 후 불편함을 느끼는 경우 로봇 보조 운동 중에 프로브를 제거하거나 느슨하게 하십시오.</li>
	<li>NIRS 측정을 완료한 후 헤드 홀더와 마킹 스티커를 제거하여 실험을 종료합니다. 참고: 장시간 착용으로 인한 두피의 피부 손상을 주의 깊게 확인하십시오.</li>
</ol>3. NIRS-SPM 소프트웨어를 이용한 질적 GLM 분석
<ol><li>MATLAB 소프트웨어에서 NIRS-SPM을 시작합니다. NIRS 장치에서 획득한 HbO2 및 HHb 농도의 변화와 관련된 데이터 파일을 NIRS-SPM 분석을 위한 파일 형식으로 변환합니다.</li>
	<li>팝업 메뉴에서 NIRS 시스템 사용 옵션을 선택합니다. 로드 버튼을 선택하고 변환 HbO2 및 HHb 농도 변경 옵션을 선택합니다.</li>
	<li>NIRS 채널 위치의 공간 등록을 검색합니다. 독립 실행형 확인란을 선택한 다음 3D 디지타이저 사용 확인란을 선택합니다.</li>
	<li>실제 좌표에서 MNI 스페이스에 이르기까지 대화 상자를 사용하여 _origin 선택합니다. 좌표 기준점 파일을 참조하고 _others CSV. 좌표 프로브/채널 파일을 참조하는 CSV입니다.</li>
	<li>등록 단추를 선택합니다. 공간 추정을 진행할 점을 선택하고 확인 버튼을 클릭합니다. 렌더링된 브레인 버튼으로 프로젝트 MNI 좌표를 클릭합니다. 참고: NIRS 채널 위치의 공간 위치는 몬트리올 신경학회(MNI) 뇌 템플릿을 기준으로 추정됩니다.</li>
	<li>등쪽 보기 옵션을 선택하고 저장 버튼을 클릭합니다.</li>
	<li>1 st 수준 지정 섹션에서 NIRS 데이터 파일 이름과 SPM 디렉터리를 선택합니다. 헤모글로빈 확인란을 선택합니다. HbO2 또는 HHb. 디자인 지정 옵션을 강조 표시하고 Sec 옵션을 선택합니다. 조건/시험 옵션 수를 강조 표시하고 숫자 7을입력합니다.</li>
	<li>시작 및 지속 시간[s] 옵션을 강조 표시하고 다음과 같이 실험 조건의 지속 시간을 곱한 개시 벡터를 입력합니다. 참고: 이 경우 발병 시간의 벡터를 [15:45:285] 또는 [15 60 105 150 195 240 285]로지정해야 합니다. 지속 시간 벡터는 [15* 것(7,1)] 또는 [15 15 15 15 15 15 15 15]로지정되어야 합니다.</li>
	<li>추세를 해제할 경우 웨이블렛-MDL 버튼을 선택합니다. 색칠 방법을 사용합니다: 로우 패스 필터와 hrf 버튼을 선택하고 직렬 상관 관계에 대해 올바른 다음 없음 단추를 선택합니다. </li>
	<li>시간적 상관 관계를 추정할 때 개별 분석을확인합니다. 참고: 개별 MNI 좌표 시스템에 fNIRS 채널 위치의 공간 지역화를 텍스트 파일로 저장합니다. 마찬가지로 개별 브로드만 영역(BA)을 기반으로 맵을 텍스트 파일로 저장합니다.</li>
	<li>시간적 상관 관계를 추정할 때 그룹 분석을확인합니다. NIRS-SPM은 MNI 표준화된 뇌 좌표 시스템에 따라 참가자 수의 평균 검안 위치를 조정했습니다.</li>
	<li>표준화된 뇌에 대한 헤모글로빈 레벨의 변화에 따라 활성화 맵을 계산합니다. HbO2 및 HHb 수준은 < 0.01의 수정되지 않은 임계값에서 중요한 것으로 간주되었습니다. 참고: 그룹 분석을 위해 오른쪽 영향을 받는 측면에서 왼쪽/오른쪽 정보가 뒤집혔습니다.</li>
</ol>4. 계층 적 혼합 모델을 기반으로 다중 채널 비교 분석
<ol><li>SAS 소프트웨어를 시작합니다. 텍스트 문서를 변환합니다(. TXT) 낮은 패스 필터로 처리된 NIRS 데이터 파일에서 HbO2 및 HHb의 농도 변화(컷오프 주파수는 0.1Hz로 설정)를 스프레드시트 소프트웨어 쉼표 분리값 파일(. CSV).</li>
	<li>프로그램을 사용하여 가져오기 SAS 데이터(.sas7bdat)를 만듭니다.</li>
	<li>다음 명령으로 가져오기 파일을 출력하고 "가져오기 파일"을 libname</li>
	<li>각 주체에 대한 사전 개입 후 파일을 출력하고 분석 SAS에서 다음 명령을 실행합니다. (그림3) 가져오기 파일을 만드는 동안, 피사체 정보를 식별할 수 있는 이름을 지정하고 개입 전(예: id1 사전, id1 게시물...)</li>
	<li>각 채널에 대한 사전 개입 및 사후 데이터 명령을 실행합니다(ch1-48; HbO2 및 HHb) 다음과 같이(그림 4).</li>
	<li>출력 결과에서 얻은 데이터를 기반으로 스프레드시트 워크북 파일(.xlsx)의 각 채널의 변경(작업 및 휴식 시 차이), 휴식 및 작업 중 값(예상 값, 상한 및 하한)의 사전 개입 후 차이(작업 및 휴식시 차이)를 입력합니다.</li>
	<li>마찬가지로 고정효과 유형 3 테스트의 상호작용 항목의 자유, F 값 및 P 값의 분자 및 분모도를 스프레드시트 통합문서 파일(.xlsx)에 입력합니다.</li>
	<li>다중 채널 테스트에서 잘못된 검색 속도(FDR)를 제어하려면 Benjamin 및 Hochberg메서드(27)를 사용하고 0.01
</ol>

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저자는 공개하는이 연구와 관련된 이해 관계의 충돌이 없습니다.

fNIRS를 위한 그룹 분석 방법에서, 질적 t-통계매핑에 의한 이미징 분석 방법을 수행하는 것 외에도, 비교멀티채널 분석을 사용하여 사전 개입(로봇 보조 운동)을 비교했습니다. 질적 분석을 위해 NIRS-SPM 소프트웨어를 일반화된 선형 모델을 기반으로 하는 대량 단변형 접근 법으로 사용했습니다. NIRS-SPM 분석은 작업 중에 활성화된 영역을 시각화하여 각 세션의 질적 결과를 보여줍니다. 더욱이...

신경 재활은 감각 운동 장애 다음 기능 회복에 중요한 역할을한다. 신경가소성 기능 회복의 메커니즘을 명확히 하기 위해 기능성 자기 공명 영상(fMRI), 양전자 방출 단층 촬영(PET), 뇌전도법(EEG), 기능적인 근적외선 분광법(fNIRS)과 같은 다양한 신경 이미징 기술이 사용되었습니다. 다른 이미징 양식에는 다른 장점과 단점이 있습니다. fMRI는 가장 일반적인 장치이지만 자기장에 의해 영향을 받고, 높은 비용, 높은 물리적 제한, 제한된 감각 운동 작업1,2,3,4. fNIRS 장치는 비침습적 광학 신경 이미징으로 두드러지며 상대적으로 낮은 공간 해상도를 가지고 있지만 fMRI4보다더 나은 시간적 해상도를 가지고 있습니다. fNIRS는 사전 개입 효과를 비교하고, 동적 모터 작업을 가지고, 휴대용, fMRI1,2,4보다자연 환경에서 더 많은 기능을 하기 때문에 치료 효과를 확인할 때 적합하다. NIRS는 뇌혈관 질환, 간질 장애, 심각한 뇌 손상, 파킨슨 병 및 인지 장애1,5의 분야에서 더 적합하다고보고되었습니다. 감각 운동 작업에 관해서는, 걸음걸이 및 서 균형6,7,8,상측 사지 기능 (손 잡기, 손가락 두드리는)8,9,복잡한 모터 스킬 트레이닝10,11,로봇 공학12,13,14,15및 뇌 컴퓨터 인터페이스16,17,18에널리 사용된다. fNIRS는 피질 대사 활동, 증가된 혈류 및 결과적으로 이차신호19로피질 활동을 측정하는 광학 신경 이미징 및 신경 혈관 커플링의 원리를 기반으로 합니다. fNIRS 신호는 혈액 산소 수준 의존fMRI20의신호와 강한 상관 관계를 가지고 보고되었다. 연속 파 fNIRS는 변형 된 맥주 램버트 법을 사용하여 분산 헤모글로빈 (HbO2)및 탈산소 화 헤모글로빈 (HHb) 피질 농도 농도 농도 의 변화를 결정하기 위해 광대역 근적외선 감쇠21,22의측정 된 변화에 따라. 연속파 NIRS 시스템을 이용하여 차동 경로 길이 계수(DPF)를 측정할 수 없기 때문에, 우리는 DPF가 일정하고 헤모글로빈 신호 변화가 밀리몰 밀리미터(mM x mm)2,18의임의 단위로 표시되었다고 가정하였다.
fNIRS 실험은 프로브 설정, 실험 설계 및 분석 방법을 포함하여 가장 적절한 방법을 선택해야 합니다. 프로브 설정에 관해서는, EEG 측정에 사용되는 국제 10-20 방법은 신경 이미징에 있는 많은 연구원에 의해 이용되는 설정 표준입니다. 최근 몇 년 동안, 몬트리올 신경 학회 (MNI) 좌표에 기초하여 표준 두뇌에 따라 좌표 설정이 사용 되었습니다. 이 실험은 일반적으로 감각 운동 작업에 사용되는 블록 설계와 이벤트 관련 설계를 사용합니다. 이것은 휴식과 작업 중에 헤모글로빈 농도의 변화를 비교하는 방법입니다. HbO2 농도 수준 증가 하 고 HHb 농도 수준 작업 의존 피 질 활동과 관련 된 대뇌 혈 류의 변화와 함께 감소. 다양한 분석 방법이 있지만 NIRS-SPM 무료 소프트웨어는 fMRI의 통계 파라메트릭 매핑(SPM)과 유사한 분석을 가능하게 합니다. NIRS 데이터의 처리는 일반 선형 모델(GLM)에 기초하여 질량 단변형 접근 방식을 사용합니다. 태스크 에 의존하는 뇌 활동 분석을 수행할 때, fNIRS 측정은 대뇌 및 초뇌구획(23)에서발동되거나 비호출되지 않은 신경 활동 및 전신 생리적 간섭(심박수, 혈압, 호흡 속도 및 자율 신경계 활동)에 의해 영향을 받을 수 있다. 따라서 사전 분석 처리, 필터링, 웨이블렛 변환 및 주 성분 분석이유용하다(23) NIRS-SPM을 이용한 데이터 처리의 필터링 및 아티팩트에 대해서는 로우패스 필터링9 및 웨이블렛 최소 설명 길이(Wavelet-MDL)24디트렌드를 사용하여 노이즈/아티팩트의 동작 또는 기타 소스를 극복하는 데 사용하였다. 이 분석 방법에 대한 자세한 내용은 Ye 외25의보고서를 참조하십시오. SPM만을 사용하는 보고서가 있지만 이미지 분석을 통해 질적 지수일 뿐이며 NIRS의 낮은 공간 해상도로 인해 그룹 분석에 주의가 필요합니다. 또한 DPF가 일정할 때 채널과 개인 간의 수치 비교를 수행해서는 안 되지만 각 채널의 변경 내용은 확인할 수 있습니다. 위의 조건에 따라 NIRS-SPM 그룹 분석 결과를 보완하기 위해 공간 등록의 정확도를 개선한 후 다중 채널 분석을 위한 원래 분석 방법을 사용했습니다. 이 멀티 채널 분석은 고정 내정간섭(전후), 고정 된 기간 (휴식 또는 작업), 무작위 개별 효과를 가진 계층 혼합 모델을 사용하여 치료 전후각 채널에서 나머지 및 작업 기간 사이의 HbO2 및 HHb 수준의 변화의 진폭을 비교했습니다.
이러한 방식으로 몇 가지 fNIRS 측정 및 분석 방법이 있습니다. 그러나 표준 메서드가 설정되지 않았습니다. 본 백서에서는 감각운동 작업을 이용한 블록 설계를 이용하여 사전 개입 의 다중 채널 fNIRS 실험에서 얻은 데이터를 분석하기 위해 질적 GLM 기반 통계 파라메트릭 매핑 및 비교 다단계 계층 적 혼합 모델을 소개합니다.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>3D-digitizer software</td>
			<td>TOPCON</td>
			<td>-</td>
			<td>NS-1000 software ver.1.50</td>
		</tr>
		<tr>
			<td>NIRS system</td>
			<td>Shimadzu</td>
			<td>-</td>
			<td>FOIRE-3000</td>
		</tr>
		<tr>
			<td>Robot</td>
			<td>CYBERDYNE</td>
			<td>-</td>
			<td>Single-joint type Hybrid Assitive Limb (HAL-SJ)</td>
		</tr>
	</tbody>
</table>

qualitative and comparative cortical activity data analyses from a functional near-infrared spectroscopy experiment applying block design

신경 이미징 연구는 재활 및 외과 치료와 같은 사전 개입 후 신경 학적 조건의 평가에 중추적 인 역할을합니다. 뇌 활동을 측정하는 데 사용되는 많은 신경 이미징 기술 중, 기능적 근적외선 분광법 (fNIRS)은 기능성 자기 공명 영상 (fMRI)과 유사한 국소 헤모글로빈 수준을 측정하여 동적 피질 활동의 평가를 가능하게합니다. 또한, fNIRS의 물리적 제한이 적기 때문에 감각 운동 작업의 여러 변종을 평가할 수 있습니다. 많은 실험실은 fNIRS 데이터 분석을 위한 몇 가지 방법을 개발했습니다. 그러나 일반적인 원칙이 동일하다는 사실에도 불구하고 보편적으로 표준화된 방법은 없습니다. 여기서는 블록 설계를 이용한 다중 채널 fNIRS 실험에서 얻은 데이터의 질적 및 비교 분석 방법을 제시합니다. 질적 분석을 위해 NIRS용 소프트웨어를 일반화된 선형 모델을 기반으로 하는 대량 단일변종 접근 법으로 사용했습니다. NIRS-SPM 분석은 작업 중에 활성화된 영역을 시각화하여 각 세션의 정성적 결과를 보여줍니다. 또한, 비침습적 3차원 디지타이저는 뇌에 비해 fNIRS 채널 위치를 추정하는 데 사용될 수 있다. NIRS-SPM 연구 결과를 확증하기 위해, 감각 운동 작업에 의해 유도된 헤모글로빈 수치의 변화의 진폭은 다중 채널 계층 적 혼합 모델을 사용하여 동일한 연구 대상의 두 개의 다른 세션 (개입 전후)에서 얻은 데이터를 비교하여 통계적으로 분석 될 수 있습니다. 우리의 방법은 운동 장애, 뇌혈관 질환 및 신경 정신 장애와 같은 다양한 신경 장애에서 사전 개입 후 분석을 측정하는 데 사용할 수 있습니다.

여기에서, 우리는 우리 그룹이 현재 일하고 있는 로봇 지원 재활을 소개합니다: 급성 뇌졸중을 가진 환자에 있는 상부 사지 모터 적자에 biofeedback 효력. 우리는 우리 병원에 입원한 10명의 동의한 뇌졸중 환자 (평균 나이: 66.8 ± 12.0 년, 2명의 여자와 8명의 남자)를 포함했습니다. 아급성 뇌졸중 단계에서, 발병 후 2 주 이상, 우리는 같은 날 상부 사지 로봇 보조 재활 전후 fNIRS ...

Watch this Scientific Journal Video about Qualitative and Comparative Cortical Activity Data Analyses from a Functional Near-Infrared Spectroscopy Experiment Applying Block Design at JoVE.com

Qualitative and Comparative Cortical Activity Data Analyses from a Functional Near-Infrared Spectroscopy Experiment Applying Block Design

우리는 감각 운동 작업을 가진 블록 디자인을 사용하여 연속 파 기능 근적외선 분광 실험의 분석을 설명합니다. 데이터 분석의 신뢰성을 높이기 위해 질적 일반 선형 모델 기반 통계 파라메트릭 매핑및 다중 채널에 대한 비교 계층 혼합 모델을 사용했습니다.

블록 설계를 적용하는 기능적인 근적외선 분광 실험에서 질적 및 비교 피질 활동 데이터 분석

Neuroimaging studies play a pivotal role in the evaluation of pre- vs. post-interventional neurological conditions such as in rehabilitation and surgical ...

qualitative-comparative-cortical-activity-data-analyses-from

Research

JoVE Journal

Neuroscience

3.6K 조회수.  Fukuoka University. 우리는 감각 운동 작업을 가진 블록 디자인을 사용하여 연속 파 기능 근적외선 분광 실험의 분석을 설명합니다. 데이터 분석의 신뢰성을 높이기 위해 질적 일반 선형 모델 기반 통계 파라메트릭 매핑및 다중 채널에 대한 비교 계층 혼합 모델을 사용했습니다.

비디오: Qualitative and Comparative Cortical Activity Data Analyses from a Functional Near-Infrared Spectroscopy Experiment Applying Block Design

Using MazeSuite and Functional Near Infrared Spectroscopy to Study Learning in Spatial Navigation

MazeSuite is a complete toolset to prepare, present and analyze navigational and spatial experiments1. MazeSuite can be used to design and edit adapted virtual 3D environments, track a participants' behavioral performance within the virtual environment and synchronize with external devices for physiological and neuroimaging measures, including electroencephalogram and eye tracking.
Functional near-infrared spectroscopy (fNIR) is an optical brain imaging technique that enables continuous, noninvasive, and portable monitoring of changes in cerebral blood oxygenation related to human brain functions2-7. Over the last decade fNIR is used to effectively monitor cognitive tasks such as attention, working memory and problem solving7-11. fNIR can be implemented in the form of a wearable and minimally intrusive device; it has the capacity to monitor brain activity in ecologically valid environments. 
Cognitive functions assessed through task performance involve patterns of brain activation of the prefrontal cortex (PFC) that vary from the initial novel task performance, after practice and during retention12. Using positron emission tomography (PET), Van Horn and colleagues found that regional cerebral blood flow was activated in the right frontal lobe during the encoding (i.e., initial na&iuml;ve performance) of spatial navigation of virtual mazes while there was little to no activation of the frontal regions after practice and during retention tests. Furthermore, the effects of contextual interference, a learning phenomenon related to organization of practice, are evident when individuals acquire multiple tasks under different practice schedules13,14. High contextual interference (random practice schedule) is created when the tasks to be learned are presented in a non-sequential, unpredictable order. Low contextual interference (blocked practice schedule) is created when the tasks to be learned are presented in a predictable order.
Our goal here is twofold: first to illustrate the experimental protocol design process and the use of MazeSuite, and second, to demonstrate the setup and deployment of the fNIR brain activity monitoring system using Cognitive Optical Brain Imaging (COBI) Studio software15. To illustrate our goals, a subsample from a study is reported to show the use of both MazeSuite and COBI Studio in a single experiment. The study involves the assessment of cognitive activity of the PFC during the acquisition and learning of computer maze tasks for blocked and random orders. Two right-handed adults (one male, one female) performed 315 acquisition, 30 retention and 20 transfer trials across four days. Design, implementation, data acquisition and analysis phases of the study were explained with the intention to provide a guideline for future studies.

MazeSuite is a complete toolset to prepare, present and analyze navigational and spatial experiments. Functional near-infrared spectroscopy (fNIR) is an optical brain imaging technique that enables noninvasive and portable monitoring of cerebral blood oxygenation changes. This paper summarizes collective use of MazeSuite and fNIR within a cognitive processing learning paradigm.

공간 탐색 구역에서 &#39;학습 연구에 적외선 분광학 가까이 MazeSuite 및 기능 사용

MazeSuite is a complete toolset to prepare, present and analyze navigational and spatial experiments1. MazeSuite can be used to design and edit adapted ...

연구

신경과학

Functional Calcium Imaging in Developing Cortical Networks

A hallmark pattern of activity in developing nervous systems is spontaneous, synchronized network activity. Synchronized activity has been observed in intact spinal cord, brainstem, retina, cortex and dissociated neuronal culture preparations. During periods of spontaneous activity, neurons depolarize to fire single or bursts of action potentials, activating many ion channels. Depolarization activates voltage-gated calcium channels on dendrites and spines that mediate calcium influx. Highly synchronized electrical activity has been measured from local neuronal networks using field electrodes. This technique enables high temporal sampling rates but lower spatial resolution due to integrated read-out of multiple neurons at one electrode. Single cell resolution of neuronal activity is possible using patch-clamp electrophysiology on single neurons to measure firing activity. However, the ability to measure from a network is limited to the number of neurons patched simultaneously, and typically is only one or two neurons. The use of calcium-dependent fluorescent indicator dyes has enabled the measurement of synchronized activity across a network of cells. This technique gives both high spatial resolution and sufficient temporal sampling to record spontaneous activity of the developing network.
A key feature of newly-forming cortical and hippocampal networks during pre- and early postnatal development is spontaneous, synchronized neuronal activity (Katz &amp; Shatz, 1996; Khaziphov &amp; Luhmann, 2006). This correlated network activity is believed to be essential for the generation of functional circuits in the developing nervous system (Spitzer, 2006). In both primate and rodent brain, early electrical and calcium network waves are observed pre- and postnatally in vivo and in vitro (Adelsberger et al., 2005; Garaschuk et al., 2000; Lamblin et al., 1999). These early activity patterns, which are known to control several developmental processes including neuronal differentiation, synaptogenesis and plasticity (Rakic &amp; Komuro, 1995; Spitzer et al., 2004) are of critical importance for the correct development and maturation of the cortical circuitry.
In this JoVE video, we demonstrate the methods used to image spontaneous activity in developing cortical networks. Calcium-sensitive indicators, such as Fura 2-AM ester diffuse across the cell membrane where intracellular esterase activity cleaves the AM esters to leave the cell-impermeant form of indicator dye. The impermeant form of indicator has carboxylic acid groups which are able to then detect and bind calcium ions intracellularly.. The fluorescence of the calcium-sensitive dye is transiently altered upon binding to calcium. Single or multi-photon imaging techniques are used to measure the change in photons being emitted from the dye, and thus indicate an alteration in intracellular calcium. Furthermore, these calcium-dependent indicators can be combined with other fluorescent markers to investigate cell types within the active network.

Spontaneous activity of developing neuronal networks can be measured using AM-ester forms of calcium-sensitive indicator dyes. Changes in intracellular calcium, indicating neuronal activation, are detected as transient changes in indicator fluorescence with one- or two-photon imaging. This protocol can be adapted for a range of developmentally-dependent neuronal networks in vitro.

두피 네트워크 개발에 기능성 칼슘 이미징

A hallmark pattern of activity in developing nervous systems is spontaneous, synchronized network activity. Synchronized activity has been observed in ...

Recording Human Electrocorticographic (ECoG) Signals for Neuroscientific Research and Real-time Functional Cortical Mapping

Neuroimaging studies of human cognitive, sensory, and motor processes are usually based on noninvasive techniques such as electroencephalography (EEG), magnetoencephalography or functional magnetic-resonance imaging. These techniques have either inherently low temporal or low spatial resolution, and suffer from low signal-to-noise ratio and/or poor high-frequency sensitivity. Thus, they are suboptimal for exploring the short-lived spatio-temporal dynamics of many of the underlying brain processes. In contrast, the invasive technique of electrocorticography (ECoG) provides brain signals that have an exceptionally high signal-to-noise ratio, less susceptibility to artifacts than EEG, and a high spatial and temporal resolution (i.e., &lt;1 cm/&lt;1 millisecond, respectively). ECoG involves measurement of electrical brain signals using electrodes that are implanted subdurally on the surface of the brain. Recent studies have shown that ECoG amplitudes in certain frequency bands carry substantial information about task-related activity, such as motor execution and planning1, auditory processing2 and visual-spatial attention3. Most of this information is captured in the high gamma range (around 70-110 Hz). Thus, gamma activity has been proposed as a robust and general indicator of local cortical function1-5. ECoG can also reveal functional connectivity and resolve finer task-related spatial-temporal dynamics, thereby advancing our understanding of large-scale cortical processes. It has especially proven useful for advancing brain-computer interfacing (BCI) technology for decoding a user's intentions to enhance or improve communication6 and control7. Nevertheless, human ECoG data are often hard to obtain because of the risks and limitations of the invasive procedures involved, and the need to record within the constraints of clinical settings. Still, clinical monitoring to localize epileptic foci offers a unique and valuable opportunity to collect human ECoG data. We describe our methods for collecting recording ECoG, and demonstrate how to use these signals for important real-time applications such as clinical mapping and brain-computer interfacing. Our example uses the BCI2000 software platform8,9 and the SIGFRIED10 method, an application for real-time mapping of brain functions. This procedure yields information that clinicians can subsequently use to guide the complex and laborious process of functional mapping by electrical stimulation.
Prerequisites and Planning:
Patients with drug-resistant partial epilepsy may be candidates for resective surgery of an epileptic focus to minimize the frequency of seizures. Prior to resection, the patients undergo monitoring using subdural electrodes for two purposes: first, to localize the epileptic focus, and second, to identify nearby critical brain areas (i.e., eloquent cortex) where resection could result in long-term functional deficits. To implant electrodes, a craniotomy is performed to open the skull. Then, electrode grids and/or strips are placed on the cortex, usually beneath the dura. A typical grid has a set of 8 x 8 platinum-iridium electrodes of 4 mm diameter (2.3 mm exposed surface) embedded in silicon with an inter-electrode distance of 1cm. A strip typically contains 4 or 6 such electrodes in a single line. The locations for these grids/strips are planned by a team of neurologists and neurosurgeons, and are based on previous EEG monitoring, on a structural MRI of the patient's brain, and on relevant factors of the patient's history. Continuous recording over a period of 5-12 days serves to localize epileptic foci, and electrical stimulation via the implanted electrodes allows clinicians to map eloquent cortex. At the end of the monitoring period, explantation of the electrodes and therapeutic resection are performed together in one procedure.
In addition to its primary clinical purpose, invasive monitoring also provides a unique opportunity to acquire human ECoG data for neuroscientific research. The decision to include a prospective patient in the research is based on the planned location of their electrodes, on the patient's performance scores on neuropsychological assessments, and on their informed consent, which is predicated on their understanding that participation in research is optional and is not related to their treatment. As with all research involving human subjects, the research protocol must be approved by the hospital's institutional review board. The decision to perform individual experimental tasks is made day-by-day, and is contingent on the patient's endurance and willingness to participate. Some or all of the experiments may be prevented by problems with the clinical state of the patient, such as post-operative facial swelling, temporary aphasia, frequent seizures, post-ictal fatigue and confusion, and more general pain or discomfort.
At the Epilepsy Monitoring Unit at Albany Medical Center in Albany, New York, clinical monitoring is implemented around the clock using a 192-channel Nihon-Kohden Neurofax monitoring system. Research recordings are made in collaboration with the Wadsworth Center of the New York State Department of Health in Albany. Signals from the ECoG electrodes are fed simultaneously to the research and the clinical systems via splitter connectors. To ensure that the clinical and research systems do not interfere with each other, the two systems typically use separate grounds. In fact, an epidural strip of electrodes is sometimes implanted to provide a ground for the clinical system. Whether research or clinical recording system, the grounding electrode is chosen to be distant from the predicted epileptic focus and from cortical areas of interest for the research. Our research system consists of eight synchronized 16-channel g.USBamp amplifier/digitizer units (g.tec, Graz, Austria). These were chosen because they are safety-rated and FDA-approved for invasive recordings, they have a very low noise-floor in the high-frequency range in which the signals of interest are found, and they come with an SDK that allows them to be integrated with custom-written research software. In order to capture the high-gamma signal accurately, we acquire signals at 1200Hz sampling rate-considerably higher than that of the typical EEG experiment or that of many clinical monitoring systems. A built-in low-pass filter automatically prevents aliasing of signals higher than the digitizer can capture. The patient's eye gaze is tracked using a monitor with a built-in Tobii T-60 eye-tracking system (Tobii Tech., Stockholm, Sweden). Additional accessories such as joystick, bluetooth Wiimote (Nintendo Co.), data-glove (5th Dimension Technologies), keyboard, microphone, headphones, or video camera are connected depending on the requirements of the particular experiment.
Data collection, stimulus presentation, synchronization with the different input/output accessories, and real-time analysis and visualization are accomplished using our BCI2000 software8,9. BCI2000 is a freely available general-purpose software system for real-time biosignal data acquisition, processing and feedback. It includes an array of pre-built modules that can be flexibly configured for many different purposes, and that can be extended by researchers' own code in C++, MATLAB or Python. BCI2000 consists of four modules that communicate with each other via a network-capable protocol: a Source module that handles the acquisition of brain signals from one of 19 different hardware systems from different manufacturers; a Signal Processing module that extracts relevant ECoG features and translates them into output signals; an Application module that delivers stimuli and feedback to the subject; and the Operator module that provides a graphical interface to the investigator.
A number of different experiments may be conducted with any given patient. The priority of experiments will be determined by the location of the particular patient's electrodes. However, we usually begin our experimentation using the SIGFRIED (SIGnal modeling For Realtime Identification and Event Detection) mapping method, which detects and displays significant task-related activity in real time. The resulting functional map allows us to further tailor subsequent experimental protocols and may also prove as a useful starting point for traditional mapping by electrocortical stimulation (ECS).
Although ECS mapping remains the gold standard for predicting the clinical outcome of resection, the process of ECS mapping is time consuming and also has other problems, such as after-discharges or seizures. Thus, a passive functional mapping technique may prove valuable in providing an initial estimate of the locus of eloquent cortex, which may then be confirmed and refined by ECS. The results from our passive SIGFRIED mapping technique have been shown to exhibit substantial concurrence with the results derived using ECS mapping10.
The protocol described in this paper establishes a general methodology for gathering human ECoG data, before proceeding to illustrate how experiments can be initiated using the BCI2000 software platform. Finally, as a specific example, we describe how to perform passive functional mapping using the BCI2000-based SIGFRIED system.

Recording Human Electrocorticographic ECoG Signals for Neuroscientific Research and Real-time Functional Cortical Mapping

We present a method for collecting electrocorticographic signals for research purposes from humans who are undergoing invasive epilepsy monitoring. We show how to use the BCI2000 software platform for data collection, signal processing and stimulus presentation. Specifically, we demonstrate SIGFRIED, a BCI2000-based tool for real-time functional brain mapping.

Neuroscientific 연구 및 실시간 기능성 두피 매핑을위한 인적 Electrocorticographic (ECoG) 신호를 녹음하기

Neuroimaging studies of human cognitive, sensory, and motor processes are usually based on noninvasive techniques such as electroencephalography (EEG), ...

Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex

Although the brain represents less than 5% of the body by mass, it utilizes approximately one quarter of the glucose used by the body at rest1. The function of non rapid eye movement sleep (NREMS), the largest portion of sleep by time, is uncertain. However, one salient feature of NREMS is a significant reduction in the rate of cerebral glucose utilization relative to wakefulness2-4. This and other findings have led to the widely held belief that sleep serves a function related to cerebral metabolism. Yet, the mechanisms underlying the reduction in cerebral glucose metabolism during NREMS remain to be elucidated.
	One phenomenon associated with NREMS that might impact cerebral metabolic rate is the occurrence of slow waves, oscillations at frequencies less than 4 Hz, in the electroencephalogram5,6. These slow waves detected at the level of the skull or cerebral cortical surface reflect the oscillations of underlying neurons between a depolarized/up state and a hyperpolarized/down state7. During the down state, cells do not undergo action potentials for intervals of up to several hundred milliseconds. Restoration of ionic concentration gradients subsequent to action potentials represents a significant metabolic load on the cell8; absence of action potentials during down states associated with NREMS may contribute to reduced metabolism relative to wake.
	Two technical challenges had to be addressed in order for this hypothetical relationship to be tested. First, it was necessary to measure cerebral glycolytic metabolism with a temporal resolution reflective of the dynamics of the cerebral EEG (that is, over seconds rather than minutes). To do so, we measured the concentration of lactate, the product of aerobic glycolysis, and therefore a readout of the rate of glucose metabolism in the brains of mice. Lactate was measured using a lactate oxidase based real time sensor embedded in the frontal cortex. The sensing mechanism consists of a platinum-iridium electrode surrounded by a layer of lactate oxidase molecules. Metabolism of lactate by lactate oxidase produces hydrogen peroxide, which produces a current in the platinum-iridium electrode. So a ramping up of cerebral glycolysis provides an increase in the concentration of substrate for lactate oxidase, which then is reflected in increased current at the sensing electrode. It was additionally necessary to measure these variables while manipulating the excitability of the cerebral cortex, in order to isolate this variable from other facets of NREMS.
	We devised an experimental system for simultaneous measurement of neuronal activity via the elecetroencephalogram, measurement of glycolytic flux via a lactate biosensor, and manipulation of cerebral cortical neuronal activity via optogenetic activation of pyramidal neurons. We have utilized this system to document the relationship between sleep-related electroencephalographic waveforms and the moment-to-moment dynamics of lactate concentration in the cerebral cortex. The protocol may be useful for any individual interested in studying, in freely behaving rodents, the relationship between neuronal activity measured at the electroencephalographic level and cellular energetics within the brain.

A procedure is described for manipulating the activity of cerebral cortical pyramidal neurons optogenetically while the electroencephalogram, electromyogram, and cerebral lactate concentration are monitored. Experimental recordings are performed on cable-tethered mice while they undergo spontaneous sleep/wake cycles. Optogenetic equipment is assembled in our laboratory; recording equipment is commercially available.

동시 Electroencephalography, 설치류 대뇌 피질의 젖산 농도와 Neuronal 활동 Optogenetic 조작의 실시간 측정

Although the brain represents less than 5% of the body  by mass, it utilizes approximately one quarter of the glucose used by the body  at rest1. The ...

Neuroimaging Field Methods Using Functional Near Infrared Spectroscopy (NIRS) Neuroimaging to Study Global Child Development: Rural Sub-Saharan Africa

Portable neuroimaging approaches provide new advances to the study of brain function and brain development with previously inaccessible populations and in remote locations. This paper shows the development of field functional Near Infrared Spectroscopy (fNIRS) imaging to the study of child language, reading, and cognitive development in a rural village setting of C&#244;te d&#39;Ivoire. Innovation in methods and the development of culturally appropriate neuroimaging protocols allow&#160;a first-time look into the brain&#39;s development and children&#39;s learning outcomes in understudied environments. This paper demonstrates protocols for transporting and setting up a mobile laboratory, discusses considerations for field versus laboratory neuroimaging, and presents a guide for developing neuroimaging consent procedures and building meaningful long-term collaborations with local government and science partners. Portable neuroimaging methods can be used to study complex child development contexts, including the impact of significant poverty and adversity on brain development. The protocol presented here has been developed for use in C&#244;te d&#39;Ivoire, the world&#39;s primary source of cocoa, and where reports of child labor in the cocoa sector are common. Yet, little is known about the impact of child labor on brain development and learning. Field neuroimaging methods have the potential to yield new insights into such urgent issues, and the development of children globally.

Neuroimaging Field Methods Using Functional Near Infrared Spectroscopy NIRS Neuroimaging to Study Global Child Development: Rural Sub-Saharan Africa

Portable neuroimaging approaches (functional Near Infrared Spectroscopy) provide advances to the study of the brain in previously inaccessible regions; here, rural C&#244;te d&#39;Ivoire. Innovation in methods and development of culturally-appropriate neuroimaging protocols permits novel study of the brain&#39;s development and children&#39;s learning outcomes in environments with significant poverty and adversity.

글로벌 아동 발달 연구에 적외선 분광학 (적외선) Neuroimaging 근처 기능 사용 하 여 Neuroimaging 필드 방법: 농촌 사하라 사막 이남의 아프리카

Portable neuroimaging approaches provide new advances to the study of brain function and brain development with previously inaccessible populations and in ...

Conducting Hyperscanning Experiments with Functional Near-Infrared Spectroscopy

Concurrent brain recordings of two or more interacting persons, an approach termed hyperscanning, are gaining increasing importance for our understanding of the neurobiological underpinnings of social interactions, and possibly interpersonal relationships. Functional near-infrared spectroscopy (fNIRS) is well suited for conducting hyperscanning experiments because it measures local hemodynamic effects with a high sampling rate and, importantly, it can be applied in natural settings, not requiring strict motion restrictions. In this article, we present a protocol for conducting fNIRS hyperscanning experiments with parent-child dyads and for analyzing brain-to-brain synchrony. Furthermore, we discuss critical issues and future directions, regarding the experimental design, spatial registration of the fNIRS channels, physiological influences and data analysis methods. The described protocol is not specific to parent-child dyads, but can be applied to a variety of different dyadic constellations, such as adult strangers, romantic partners or siblings. To conclude, fNIRS hyperscanning has the potential to yield new insights into the dynamics of the ongoing social interaction, which possibly go beyond what can be studied by examining the activities of individual brains.

The present protocol describes how to conduct fNIRS hyperscanning experiments and analyze brain-to-brain synchrony. Further, we discuss challenges and possible solutions.

Hyperscanning 기능성 근 적외선 분광학 실험 실시

Concurrent brain recordings of two or more interacting persons, an approach termed hyperscanning, are gaining increasing importance for our understanding ...

Conducting Concurrent Electroencephalography and Functional Near-Infrared Spectroscopy Recordings with a Flanker Task

Concurrent EEG and fNIRS recordings offer an excellent opportunity to gain a full understanding of the neural mechanism of cognitive processing by inspecting the relationship between the neural and hemodynamic signals. EEG is an electrophysiological technology that can measure the rapid neuronal activity of the cortex, whereas fNIRS relies on the hemodynamic responses to infer brain activation. The combination of EEG and fNIRS neuroimaging techniques can identify more features and reveal more information associated with the functioning of the brain. In this protocol, fused EEG-fNIRS measurements were performed for concurrent recordings of evoked-electrical potentials and hemodynamic responses during a Flanker task. In addition, the critical steps for setting up the hardware and software system as well as the procedures for data acquisition and analysis were provided and discussed in detail. It is expected that the present protocol can pave a new avenue for improving the understanding of the neural mechanisms underlying various cognitive processes by using the EEG and fNIRS signals.

The present protocol describes how to perform concurrent EEG and fNIRS recordings and how to inspect the relationship between the EEG and fNIRS data.&#160;

플랭커 태스크를 이용한 동시 전류 뇌파 및 기능적 근적외선 분광 기록 수행

Concurrent EEG and fNIRS recordings offer an excellent opportunity to gain a full understanding of the neural mechanism of cognitive processing by ...

Combined In Vivo Anatomical and Functional Tracing of Ventral Tegmental Area Glutamate Terminals in the Hippocampus

Optogenetic modulation of neuron sub-populations in the brain has allowed researchers to dissect neural circuits in vivo and ex vivo. This provides a premise for determining the role of neuron types within a neural circuit, and their significance in information encoding relative to learning. Likewise, the method can be used to test the physiological significance of two or more connected brain regions in awake and anesthetized animals. The current study demonstrates how VTA glutamate neurons modulate the firing rate of putative pyramidal neurons in the CA1 (hippocampus) of anesthetized mice. This protocol employs adeno-associated virus (AAV)-dependent labeling of VTA glutamate neurons for the tracing of VTA presynaptic glutamate terminals in the layers of the hippocampus. Expression of light-controlled opsin (channelrhodopsin; hChR2) and fluorescence protein (eYFP) harbored by the AAV vector permitted anterograde tracing of VTA glutamate terminals, and photostimulation of VTA glutamate neuron cell bodies (in the VTA). High-impedance acute silicon electrodes were positioned in the CA1 to detect multi-unit and single-unit responses to VTA photostimulation in vivo. The results of this study demonstrate the&#160;layer-dependent distribution of presynaptic VTA glutamate terminals in the hippocampus (CA1, CA3, and DG). Also, the photostimulation of VTA glutamate neurons increased the firing and burst rate of putative CA1 pyramidal units in vivo.

The current protocol demonstrates a simple method for tracing of ventral tegmental area (VTA) glutamate projections to the hippocampus. Photostimulation of VTA glutamate neurons was combined with CA1 recording to demonstrate how VTA glutamate terminals modulate CA1 putative pyramidal firing rate in vivo.

해마의 생체 내 해부학 및 기능적 추적

Optogenetic modulation of neuron sub-populations in the brain has allowed researchers to dissect neural circuits in vivo and ex vivo. This provides a ...

Analyses of Actin Dynamics, Clutch Coupling and Traction Force for Growth Cone Advance

To establish functional networks, neurons must migrate to their appropriate destinations and then extend axons toward their target cells. These processes depend on the advances of growth cones that located at the tips of neurites. Axonal growth cones generate driving forces by sensing their local microenvironment and modulating cytoskeletal dynamics and actin-adhesion coupling (clutch coupling). Decades of research have led to the identification of guidance molecules, their receptors, and downstream signaling cascades for regulating neuronal migration and axonal guidance; however, the molecular machineries required for generating forces to drive growth cone advance and navigation are just beginning to be elucidated. At the leading edge of neuronal growth cones, actin filaments undergo retrograde flow, which is powered by actin polymerization and actomyosin contraction. A clutch coupling between F-actin retrograde flow and adhesive substrate generates traction forces for growth cone advance. The present study describes a detailed protocol for monitoring F-actin retrograde flow by single speckle imaging. Importantly, when combined with an F-actin marker Lifeact, this technique can quantify 1) the F-actin polymerization rate and 2) the clutch coupling efficiency between F-actin retrograde flow and the adhesive substrate. Both are critical variables for generating forces for growth cone advance and navigation. In addition, the present study describes a detailed protocol of traction force microscopy, which can quantify 3) traction force generated by growth cones. Thus, by coupling the analyses of&#160;single speckle imaging and traction force microscopy, investigators can monitor the molecular mechanics underlying growth cone advance and navigation.

To advance, growth cones must exert traction forces against the external environment. The generation of traction forces is dependent on actin dynamics and clutch coupling. The present study describes methods for analyzing actin dynamics, clutch coupling and traction forces for growth cone advance.

액틴 다이내믹스, 클러치 커플링, 성장 콘 발전을 위한 견인력 분석

To establish functional networks, neurons must migrate to their appropriate destinations and then extend axons toward their target cells. These processes ...

Group Synchronization During Collaborative Drawing Using Functional Near-Infrared Spectroscopy

Functional near-infrared spectroscopy (fNIRS) is a noninvasive method particularly suitable for measuring cerebral cortex activation in multiple subjects, which is relevant for studying group interpersonal interactions in ecological settings. Although many fNIRS systems technically offer the possibility to monitor more than two individuals simultaneously, establishing easy-to-implement setup procedures and reliable paradigms to track hemodynamic and behavioral responses in group interaction is still required. The present protocol combines fNIRS and video-based observation to measure interpersonal synchronization in quartets during a cooperative task.This protocol provides practical recommendations for data acquisition and paradigm design, as well as guiding principles for an illustrative data analysis example. The procedure is designed to assess differences in brain and behavior interpersonal responses between social and non-social conditions inspired by a well-known ice-breaker activity, the Collaborative Face Drawing Task. The described procedures can guide future studies to adapt group naturalistic social interaction activities to the fNIRS environment.

The present protocol combines functional near-infrared spectroscopy (fNIRS) and video-based observationto measure interpersonal synchronization in quartets during a collaborative drawing task.

기능적 근적외선 분광법을 사용한 협업 드로잉 중 그룹 동기화

Functional near-infrared spectroscopy (fNIRS) is a noninvasive method particularly suitable for measuring cerebral cortex activation in multiple subjects, ...