이 작품은 신경 통신 기술을위한 센터 (CNCT), 바이오 메디컬 이미징 및 생물의 국립 연구소 (NIBIB, P41 EB002030)에 의해 재정 지원과 국립 보건원 (NIH)에서 지원하는 P41 자원 센터에 의해 재정 지원되었다. 저자들은 수술 기법의 발전을 위해 NeuroNexus 기술의 리오 Vetter 감사하고자, 그리고 수술시 사용되는 장비의 목록을 컴파일하는 동안 미시간 대학의 파텔을 Paras.

1. 사전 주입 절차 (공통) <ol><li> 에 잘 귀 뒤의 두 눈 사이에서 동물의 머리의 정상을 면도. 머리를 면도하는 이발사의 전기 면도기를 사용합니다. 다음, 이소 프로필 알코올 처음에 머리를 빼주 betadine (3 배 반복). 안구 윤활제를 적용합니다. </li><li> stereotaxic 귀에 막대에서 동물을 탑재합니다. 운하에 대해 머리를 옆으로 이동하여 동물의 권리 (内耳)의 유스 타키 오관으로 오른쪽 stereotaxic 귀 표시줄의 고정 팁을 직접. 익스 트림 치료는 각 귀 막대 끝이 제대로 (外耳道)에 위치되어 있는지 확실히하기 위해 이동해야합니다. 다음으로 유스 타키 오관으로 느슨하게 왼쪽 귀 막대를 푸시하고 ​​약간 안쪽으로 압력을 적용합니다. 동물의 코를 잡고 앞뒤로 단단히 그것을 흔드는. 동물이 마운트되어 있으면 제대로 머리 rigidly 탑재 느낄 것이다. </li></ol> 2. 사전 주입 수술 (공용) <ol><li> 미세 가위로 두 개의 절반 유통 잘라레 중간선 바깥쪽에서 부분적으로 두피를 제거 </li><li> 뼈에 부합 골막 결합 조직을 다쳤어요하​​고 제거합니다. 이러한 메스 블레이드 또는 멸균 면봉 끝의 뒷면과 같은 뭉툭한 가장자리로된다고하는 것은 모세 혈관이 깔린보다는 부드럽게 절단되기 때문에 출혈이 최소화됩니다. 제어 뼈 출혈 뼈 왁스 또는 cauterizing 아이언을 사용합니다. </li><li> 소독 및 과산화수소를 사용하여 뼈 표면을 청소하십시오. 더 명확하게 보이는 두개골 봉합를 만들기 위해 뼈를 건조합니다. Bregma 및 람다 포인트는 언급하고 머리를 날려하는 데 사용해야합니다. craniotomy 및 전극 이식을위한 위치 Bregma과 관련하여 측정해야하며 일시적으로 볼펜이나 표면에 shallowly 드릴링에 의해 표시됩니다. craniotomy 위치는 연구의 실험 목적 (예 : 모터, 관능 실험 1)에 따라 달라집니다. </li><li> 뼈로 screwholes를 파악 및 지상 배치 및 s를 안정화대원. 항상 스테인레스 스틸 또는 티타늄 나사를 사용합니다. 가능한 한 인치당 많은 스레드가 바람직한만큼 소규모 보석의 나사 (예 : # 2-56, 또는 # 00-80 용). </li></ol> 3. 실리콘 전극 임플란트 절차 <ol><li> 나사를 이온 주입 후 craniotomy를 드릴 다운하십시오. craniotomy의 모서리 이미 네 얕은 시험 점수를 드릴 다운하십시오. 가이드 craniotomy의 경계를 드릴로 파일럿 부호를 사용합니다. 튼튼한 포셉 한 켤레를 사용 뚫고되지 뼈의 중심 질량을 제거합니다. 오픈 craniotomy의 가장자리는 추가 시추로 청소해야 할 수도 있습니다. 골의 얇은 조각은 microrongeurs 한 켤레로 이어 수 있습니다. 이러한 중간선에 세로 봉합 1 0.5 mm 이내에 나타날 수 있습니다 화살 부비동 등 주요 혈관을 피하기 위해 신중하게 수술을 계획합니다. 화살 부비동을 펑쳐링하는 것은 광범위한 출혈을 일으킬 수 있습니다. 노출 경질은 aCSF이나 식염수가 충분 보관해야합니다. </li><li> 문양의 경질의 mater으로당신은 패션 작은 게이지의 멸균 피하 주사를 (예를 들어, &lt;28 게이지)를 사용하여 작은 후크를해야합니다. 바늘의 베벨를 타고 지점에서 90도 굽힘을 형성하는 약간 틸팅 (예 : 메스의 평평한 부분 등) 확고한 표면에 대해 그것이 평평한 누르십시오. 경질의 표면을 잡으면, 그리고 뇌 표면에서 위쪽으로 들어올리이 훅을 사용하십시오. 경질은 마이크로 가위를 사용하여자를 수 또는 경질의 후크의 수평적 움직임을 사용하여 찢어지지 수 있습니다. 경질 후크를 사용할 때 두뇌의 표면에 혈관을 피하기 위해주의를 기울입니다. 당신은 CSF는 경질이 resected 때 밖으로 누출과 경질의 표면으로부터 색상의 차이가주의해야합니다. </li><li> 조심스럽게 그라운드 접지 나사의 기지 주변의 실리콘 전극 와이어 및 치과 시멘트와 장소에서 전술을 첨부합니다. 실리콘 전극은 전극 케이블 리드가 craniotomy 위에 아치 형의 것을 주의깊게 있도록 배치해야합니다. </li><li> 임플란트가 리니어 모터 액추에이터 괞찮아를 사용하여 배치됩니다stereotaxic 장치에 ced. 삽입 장치에 전극을 부착하려면, 폴리에틸렌 글리콜 (PEG)은 납땜 인두로 가열하고 삽입 막대와 기록 전극에 적용됩니다. </li><li> 전극은 확보를 통해 수동으로 두뇌의 표면에 원하는 stereotaxic 위치로 전극의 끝부분을 낮춥니다. </li><li> 선형 액추에이터에 소프트웨어 인터페이스를 사용하여 뇌 조직 내에서 원하는 깊이에 전극을 이동합니다. </li><li> 전극 케이블에 인접한 나사에서 PEG를 부착하여 두뇌에 기록 전극을 고정합니다. 이제 구속 PEG를 해산하기 위해 염분을 추가하여 삽입 장치에서 전극을 제거할 수 있습니다. 멀리 각 전극에 대한 craniotomy 및 반복에서 삽입 장치를 올립니다. </li><li> 식염수 젖은 Gelfoam으로 노출 craniotomy를 커버. 실리콘 폴리머는 다음 전극 케이블을 차지하고 있으며 치과 아크릴로부터 보호하는 데 사용됩니다. 마지막에있는 커넥터 위치위치는 다음 강력한 headcap를 만들기 위해 치과 아크릴을 적용합니다. </li></ol> 4. 전극 임플란트 절차 MicroECoG <ol><li> microECoG 전극의 주입은 약간 큰 craniotomy를 포함합니다. 5 × 5 밀리미터 장치의 경우 6 X 6mm의 craniotomy을하셔야합니다. 아직 경질 또는 PIA를 만지고 위험에 건조하고없는 상태에서 craniotomy을 뚫는 전에 UV-치료 가능한 치과 아크릴은 craniotomy 사이트의 주변에 적용됩니다. </li><li> 멸균 수술 훈련은 craniotomy의 일반적인 모양의 뼈 표면을 자세히 보려면​​ # 107 버는 함께 사용됩니다. 작고 # 106 버는 그런 다음 얇은 투명 레이어로 골의 나머지 부분을 아래로 밖으로 드릴 수 있습니다. Rongeurs 다음 뼈의 나머지 조각을 날아갈 수있을 것입니다. microECoG는 epidurally 또는 subdurally 이식 수 있습니다. 다시 말하지만, 인공 CSF 또는 식염수 경질 잘 충분하게. </li><li> microECoG을 이식하기 위해서는 ope 위에 stereotaxic 팔을 배치N 개의 두개골과 무균 테이프를 사용하여 팔을에 전극을 고정하십시오. 전극은 지금 craniotomy로 아래로 낮아 수 있습니다. 전극의 사이트가 아래로 향하게되고 경질 또는 pial 표면과 접촉하는 것입니다 있는지 확인합니다. 습한 보관하면 전극이 조직에 밀어해야하며 자리에 남아있을 것입니다. </li><li> 접지 와이어는 자체가 이상 이하 적어도 세 번 주위에 배치하여 접지 나사에 연결됩니다. 접지 나사는 경질와 접촉으로 온 모든 나사가 될 수 있습니다. 참조 와이어도 같은 방식으로 참조 나사에 묶여 수 있습니다. </li><li> 경질 또는 PIA가 노출이 어디 식염수 젖은 Gelfoam의 작은 조각은 전극을 둘러싼 배치해야합니다. 식염수 젖은 Gelfoam의 소량 또한 박막 전극의 상단을 커버한다. UV-치료 가능한 치과 아크릴는 Gelfoam의 상단에 적용할 수 있으며, 안정적인 헤드 캡을 만드는 데 사용할 수 있습니다. 아크릴은 박막 케이블 coveri에 직접 적용NG 그것 커넥터에 도달할 때까지. 커넥터 덮개의 하단 넘어 커버하지 않도록주의하십시오. </li><li> 또는 4.3에서 microECoG의 주입은 즉시 microECoG 기판에 제조 작은 구멍을 통해 실리콘 microelectrode 배열 (단계 3.2-3.6)의 주입 다음에 할 수 있습니다. 이것은 주위의 뇌 표면의 고해상도 필드 레코딩과 함께 다양한 깊이에서 단일 단위의 동시 녹음을 허용합니다. </li></ol> 5. 수술후의 회복 <ol><li> 시멘트가 완전히 가득 찼어 후, 헤드 캡 주위에 단단히 피부를 치료하고 stereotaxic 프레임에서 동물을 제거합니다. </li><li> copiously 상처 주위 살균 분말 또는 항생 연고를 바릅니다. 귀를으로부터 출혈이있다면, 외이도에 일부 항생제 잘 넣으십시오. </li><li> barbiturates는 체온을 유지로부터 동물을 막기로 동물이 따뜻하게있어. 하우스 EACH는 만성 별도의 새장에 동물을 이식. 그것은 보통 수술로 완전히 복구하는 동물을위한 4-7일 소요됩니다. </li></ol> 6. 대표 결과 성공적인 실리콘 전극 이식 수술 500kOhm-2MOhm 사이에 측정 임피던스와 함께 사이트를 녹화하고, 10kOhm-50kOhm (1kHz에서) 사이에 박막 전극됩니다. 신경 신호뿐만 아니라 즉시 사후 수술을 확인하실 수 있습니다. 당신은 이식 전극에 스파이크를보고 박막 표면 전극 (그림 1)에서 느린 웨이브 oscillations을 볼 수 있어야합니다. <img src="/files/ftp_upload/3565/3565fig1.jpg" alt="그림 1" /> 1 그림. 하나의 유닛 활동과 ECoG의 oscillations의 대표 결과입니다. 네크로 텍스에 이식 만성 실리콘 전극 6 동시에 기록된 채널의) 와이드 밴드가 데이터입니다. 스파이크가 자주 녹화 사이트를 통해 볼 수 있습니다. B) E두뇌의 표면에 16 채널 박막 uECoG 전극 배열의 6 채널의 코그의 oscillations. 16 개 신호의 평균은 각 성분 (일반 평균 참조)​​에서 제거되었으며, 500 Hz에서 디지털 로우 패스 필터가 적용되었다.

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DRK는 NeuroNexus 기술의 일부 소유자입니다.

4,5 neuroprosthetic 체제 microstimulation 3 또는 제어 신호를 제공, 뇌 기능 2 연구에 대한 intracortical 및 표면 다채널 녹화 인터페이스를 사용하는 점점 관심이 있습니다. 이 비디오에 제시된 방법은 만성 관통 및 표면 전극 시스템을 이식하는 방법을 보여줍니다. 다른 실리콘 만성 전극 시스템은 6 존재하는 동안, 우리는 NeuroNexus 테크놀로지 7,8 개발한 평면 전극을 이온 주입에 주력했습니다. 다른 전극 시스템의 이식을위한 기술은 상당히 다릅니다. 설명 외과 단계, stereotaxic 귀 클럽에서 동물을 장착하는 것이 가장 어렵고 이식 전극의 배치의 정확성을 결정하는 가장 중요한 단계가 될 수 있습니다. 익스 트림 치료는 각 귀 막대 끝이 제대로 (外耳道)에 위치하고있다는 생각이 따라서 이동해야합니다. assura 후왼손 엄지와 집게손가락이 고개를 지원하고 귀, 바 팁 반대 머리의 압력을 계속 적용하면서 귀 바 팁은 동물의 권리 귀 (外耳道)에 있는지 제정신, 네 오른손으로 동물의 목을 놓습니다. 오른손으로 유스 타키 오관으로 느슨하게 왼쪽 귀 막대를 푸시하고 ​​약간 안쪽으로 압력을 적용합니다. 당신이 (外耳道)로 왼쪽 귀, 바 팁 전표를 느낄 수있을 때까지 아래로 앞으로 모든 주변 동물의 머리의 왼쪽으로 이동합니다. 오른손으로 왼쪽 귀 표시줄에 압력을 적용하는 것을 계속한다. 동물의 머리를 풀어하며, 여러분 이제 무료로 왼손으로, 왼쪽 귀 표시줄의 세트 나사를 조입니다.

<table border="1"><tbody><tr><td> 장비 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> 뼈 스크류 - # 2-56 X 16분의 3, 필립스 냄비 머리, 18-8 S / S </td><td> BoltDepot </td><td> 7650 </td><td></td></tr><tr><td> 뼈 스크류 - # 2-56 × 1 / 8, 필립스 냄비 머리, 18-8 S / S </td><td> BoltDepot </td><td> 9617 </td><td></td></tr><tr><td> 대형 산소 탱크 - 크기 K </td><td> 극저온 가스 </td><td> OXKME </td><td></td></tr><tr><td> 작은 산소 탱크 - 사이즈 전자 </td><td> 극저온 가스 </td><td> OXYE-AL </td><td></td></tr><tr><td> Vannas - 튀빙겐 봄 가위 - 2.5mm 블레이드 각도 </td><td> 파인 과학 도구 </td><td> 15002-08 </td><td></td></tr><tr><td> 뒤몽 # 5 포셉, 테플론 </td><td> 파인 과학 도구 </td><td> 11626-11 </td><td></td></tr><tr><td> 뒤몽 # 5 포셉, Dumostar </td><td> 파인 과학 도구 </td><td> 11295-10 </td><td></td></tr><tr><td> 학생 Vannas 봄 가위 </td><td> 파인 과학 도구 </td><td> 91500-09 </td><td></td></tr><tr><td> Adson 집게 </td><td> 파인 과학 도구 </td><td> 11006-12 </td><td></td></tr><tr><td> 구부러져 Friendman - 피어슨 Ronqeur, </td><td> 파인 과학 도구 </td><td> 16221-14 </td><td></td></tr><tr><td> 아이리스 가위 - ToughCut </td><td> 파인 과학 도구 </td><td> 14058-11 </td><td></td></tr><tr><td> Vannas - 튀빙겐 봄 가위 - 5mm 블레이드 스트레이트 </td><td> Fi를NE 과학 도구 </td><td> 15003-08 </td><td></td></tr><tr><td> 메스 핸들 </td><td> 파인 과학 도구 </td><td> 10003-12 </td><td></td></tr><tr><td> 플라스틱 살균 상자 </td><td> 파인 과학 도구 </td><td> 20810-02 </td><td></td></tr><tr><td> 스테인레스 스틸 눈금자 </td><td> 파인 과학 도구 </td><td> 30086-15 </td><td></td></tr><tr><td> 프로브 및 주걱 - 스트레이트 14cm </td><td> 파인 과학 도구 </td><td> 10090-13 </td><td></td></tr><tr><td> Hemostat 스트레이트 </td><td> 파인 과학 도구 </td><td> 13002-10 </td><td></td></tr><tr><td> Hemostat가 구부러져 </td><td> 파인 과학 도구 </td><td> 13003-10 </td><td></td></tr><tr valign="bottom"><td> 마이크로 드릴 역inless 철강 털은 - 0.7mm </td><td> 파인 과학 도구 </td><td> 19008-07 </td><td> 10/Pack </td></tr><tr valign="bottom"><td> 마이크로 훈련 스테인레스 철강 털은 - 0.5mm </td><td> 파인 과학 도구 </td><td> 19007-05 </td><td> 10/Pack </td></tr><tr valign="bottom"><td> 셀프 부수 뼈 나사 - 1.17mm (diam) - 4.7mm (길이) </td><td> 파인 과학 도구 </td><td> 19010-00 </td><td> 100/Pack </td></tr><tr valign="bottom"><td> 인공 눈물 </td><td> 어부 </td><td> NC9860842 </td><td> 각각의 </td></tr><tr valign="bottom"><td> Betadine </td><td> 어부 </td><td> 19-066452 </td><td> 병 </td></tr><tr valign="bottom"><td> F-AIR 카본 필터 </td><td> 어부 </td><td> NC9112250 </td><td> 깡통 </td></tr><tr valign="bottom"><td> 작은 주걱 면화는 끌어내려 &#39;무균 비 </td> &lt;TD&gt; Materiel 서비스 </td><td> 1104 </td><td> 10Pack/Case </td></tr><tr valign="bottom"><td> 거즈 2 &quot;X 2&quot; </td><td> Materiel 서비스 </td><td> 1630 </td><td> 25Pack/Case </td></tr><tr valign="bottom"><td> 니들 팁, 23 GA X 1 &quot; </td><td> Materiel 서비스 </td><td> 39,412 </td><td> 케이스가 판매 </td></tr><tr valign="bottom"><td> 니들 팁 (27) GA X 1.25 &quot; </td><td> Materiel 서비스 </td><td> 25,251 </td><td></td></tr><tr valign="bottom"><td> 니들 팁, 30 GA X 0.5 &quot; </td><td> Materiel 서비스 </td><td> 22,023 </td><td> 10Box/Case </td></tr><tr valign="bottom"><td> 연고, 트리플 항생제 0.6g </td><td> Materiel 서비스 </td><td> 2528 </td><td></td></tr><tr valign="bottom"><td> 파우치, 셀프 인감 살균 Pouche<html> 초, 3.5 &quot;X 8&quot; </td><td> Materiel 서비스 </td><td> 1023 </td><td> 10Box/Case </td></tr><tr valign="bottom"><td> 솔루션 1000ml를 Ringers </td><td> Materiel 서비스 </td><td> 5263 </td><td> 12Bag/Case </td></tr><tr valign="bottom"><td> 메스 블레이드 # 15 멸균 일회용 </td><td> Materiel 서비스 </td><td> 1975 </td><td> 3Box/Case </td></tr><tr valign="bottom"><td> 봉합사, 나일론 w/P-3 니들, 5-0, 18인치 </td><td> Materiel 서비스 </td><td> 4618 </td><td> 12/Box </td></tr><tr valign="bottom"><td> Underpad, 일회용 23 &quot;X 36&quot; </td><td> Materiel 서비스 </td><td> 2545 </td><td> 15Pack/Case </td></tr><tr valign="bottom"><td> 윤활제, 젤리 외과 무균 </td><td> Materiel 서비스 </td><td> 2538 </td><td> 12Box/Case </td></tr>&quot;&gt; <td> 흡수성 폼 젤 2cmx6cmx7mm </td><td> Materiel 서비스 </td><td> 3107 </td><td></td></tr><tr valign="bottom"><td> 여섯 조각 세트 - 나사 스크루 드라이버 세트 </td><td> 국립 보석점 용품 </td><td> ETSCR98001 </td><td></td></tr><tr valign="bottom"><td> Kwik 실, 접착제 </td><td> WPI </td><td> KWIK-SIL </td><td></td></tr><tr valign="bottom"><td> Kwik 실 튜브 </td><td> WPI </td><td> 600,009 </td><td></td></tr><tr valign="top"><td> 만성 실리콘 전극 16-64 사이트 </td><td> NeuroNexus 기술 </td><td> 프로브 디자인에 다름 </td><td></td></tr><tr valign="top"><td> 만성 박막 표면 전극 (microECoG) 16-32 사이트 </td><td> NeuroNexus 기술 </td><td> 프로브 디자인에 다름 </td><td>&nbsp;
 </td>
 </tr>
 </tbody>
</table>

surgical implantation of chronic neural electrodes for recording single unit activity and electrocorticographic signals

장기 electrophysiological 녹음의 성공은 종종 주입 수술의 품질에 따라 달라집니다. 여기 전극 시스템을 이온 주입하는 과정을 배우고있는 외과의에 대한 유용한 정보를 제공합니다. 우리는 관통 및 표면 전극 모두의 주입 절차를 보여줍니다. 수술 과정은 절차에 걸쳐 각 단계의 자세한 설명을 포함하여 처음부터 끝까지 설명되어 있습니다. 그것은이 비디오 가이드 설치류 모델 및 기타 소형 동물 모델에서 실시 절차를 향해 초점을 맞추고있다는 또한 지적되어야한다. 설명된 절차의 수정은 다른 동물 모델에 가능합니다.

Watch this Scientific Journal Video about Surgical Implantation of Chronic Neural Electrodes for Recording Single Unit Activity and Electrocorticographic Signals at JoVE.com

Surgical Implantation of Chronic Neural Electrodes for Recording Single Unit Activity and Electrocorticographic Signals

우리는 만성 신경 기록 전극을 이온 주입하는 과정을 배우고있는 외과의에 대한 유용한 정보를 제공합니다. 관통 및 표면 양쪽 전극 시스템을위한 기술은 쥐 동물 모델에 설명되어 있습니다.

단일 유닛 활동과 Electrocorticographic 신호를 녹음하기 위해 만성 신경 전극 외과 이식

The success of long-term electrophysiological recordings often depends on the quality of the implantation surgery.  Here we provide useful information for ...

surgical-implantation-chronic-neural-electrodes-for-recording-single

Research

JoVE Journal

Neuroscience

47.1K 조회수.  University of Michigan. 우리는 만성 신경 기록 전극을 이온 주입하는 과정을 배우고있는 외과의에 대한 유용한 정보를 제공합니다. 관통 및 표면 양쪽 전극 시스템을위한 기술은 쥐 동물 모델에 설명되어 있습니다.

비디오: Surgical Implantation of Chronic Neural Electrodes for Recording Single Unit Activity and Electrocorticographic Signals

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), ...

연구

신경과학

Optical Recording of Suprathreshold Neural Activity with Single-cell and Single-spike Resolution

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also propagation of suprathreshold spiking activity involves populations of neurons. Empirical studies addressing cortical function directly thus require recordings from populations of neurons with high resolution. Here we describe an optical method and a deconvolution algorithm to record neural activity from up to 100 neurons with single-cell and single-spike resolution. This method relies on detection of the transient increases in intracellular somatic calcium concentration associated with suprathreshold electrical spikes (action potentials) in cortical neurons. High temporal resolution of the optical recordings is achieved by a fast random-access scanning technique using acousto-optical deflectors (AODs)1. Two-photon excitation of the calcium-sensitive dye results in high spatial resolution in opaque brain tissue2. Reconstruction of spikes from the fluorescence calcium recordings is achieved by a maximum-likelihood method. Simultaneous electrophysiological and optical recordings indicate that our method reliably detects spikes (&gt;97% spike detection efficiency), has a low rate of false positive spike detection (&lt; 0.003 spikes/sec), and a high temporal precision (about 3 msec) 3. This optical method of spike detection can be used to record neural activity in vitro and in anesthetized animals in vivo3,4.

Understanding the function of the vertebrate central nervous system requires recordings from many neurons because cortical function arises on the level of populations of neurons. Here we describe an optical method to record suprathreshold neural activity with single-cell and single-spike resolution, dithered random-access scanning. This method records somatic fluorescence calcium signals from up to 100 neurons with high temporal resolution. A maximum-likelihood algorithm deconvolves the underlying suprathreshold neural activity from the somatic fluorescence calcium signals. This method reliably detects spikes with high detection efficiency and a low rate of false positives and can be used to study neural populations in vitro and in vivo.

단일 셀 및 단일 스파이크 해상도 suprathreshold 신경 활동의 광 기록

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

메뚜기에 신경 활동을 특성화하기 위해 다중 단위 기록 방법 (<em&gt; Schistocerca 아메리카나</em&gt;) 강한 회로

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

A Method for Systematic Electrochemical and Electrophysiological Evaluation of Neural Recording Electrodes

New materials and designs for neural implants are typically tested separately, with a demonstration of performance but without reference to other implant characteristics.&#160;This precludes a rational selection of a particular implant as optimal for a particular application and the development of new materials based on the most critical performance parameters.&#160;This article develops a protocol for in vitro and in vivo testing of neural recording electrodes.&#160;Recommended parameters for electrochemical and electrophysiological testing are documented with the key steps and potential issues discussed.&#160;This method eliminates or reduces the impact of many systematic errors present in simpler in vivo testing paradigms, especially variations in electrode/neuron distance and between animal models.&#160;The result is a strong correlation between the critical in vitro and in vivo responses, such as impedance and signal-to-noise ratio.&#160;This protocol can easily be adapted to test other electrode materials and designs.&#160;The in vitro techniques can be expanded to any other nondestructive method to determine further important performance indicators.&#160;The principles used for the surgical approach in the auditory pathway can also be modified to other neural regions or tissue.

Different electrode coatings affect neural recording performance through changes to electrochemical, chemical and mechanical properties. Comparison of electrodes in vitro is relatively simple, however comparison of in vivo response is typically complicated by variations in electrode/neuron distance and between animals. This article provides a robust method to compare neural recording electrodes.

신경 기록 전극의 체계적인 전기 및 전기 생리 평가를위한 방법

New materials and designs for neural implants are typically tested separately, with a demonstration of performance but without reference to other implant ...

Extracellular Recording of Neuronal Activity Combined with Microiontophoretic Application of Neuroactive Substances in Awake Mice

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and awake animals. Therefore, methods allowing the manipulation of synaptic systems in awake animals are required in order to determine the contribution of synaptic inputs to neuronal processing unaffected by anesthetics. Here, we present methodology for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. By combining these procedures, we performed microiontophoretic injections of gabazine to selectively block GABAA receptors in neurons of the inferior colliculus of head-restrained mice. Gabazine successfully modified neural response properties such as the frequency response area and stimulus-specific adaptation. Thus, we demonstrate that our methods are suitable for recording single-unit activity and for dissecting the role of specific neurotransmitter receptors in auditory processing.
The main limitation of the described procedure is the relatively short recording time (~3 hr), which is determined by the level of habituation of the animal to the recording sessions. On the other hand, multiple recording sessions can be performed in the same animal. The advantage of this technique over other experimental procedures used to manipulate the level of neurotransmission or neuromodulation (such as systemic injections or the use of optogenetic models), is that the drug effect is confined to the local synaptic inputs to the target neuron. In addition, the custom-manufacture of electrodes allows adjustment of specific parameters according to the neural structure and type of neuron of interest (such as the tip resistance for improving the signal-to-noise ratio of the recordings).

We present methods for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. This technique allows the detailed analysis of putative local synaptic inputs to the neuron of interest.

깨어 마우스의 신경 활성 물질의 Microiontophoretic 응용 프로그램과 결합하여 신경 활동의 세포 외 기록

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and ...

Fabrication of High Contact-Density, Flat-Interface Nerve Electrodes for Recording and Stimulation Applications

Many attempts have been made to manufacture multi-contact nerve cuff electrodes that are safe, robust and reliable for long term neuroprosthetic applications. This protocol describes a fabrication technique of a modified cylindrical nerve cuff electrode to meet these criteria. Minimum computer-aided design and manufacturing (CAD and CAM) skills are necessary to consistently produce cuffs with high precision (contact placement 0.51 &#177; 0.04 mm) and various cuff sizes. The precision in spatially distributing the contacts and the ability to retain a predefined geometry accomplished with this design are two criteria essential to optimize the cuff&#39;s interface for selective recording and stimulation. The presented design also maximizes the flexibility in the longitudinal direction while maintaining sufficient rigidity in the transverse direction to reshape the nerve by using materials with different elasticities. The expansion of the cuff&#39;s cross sectional area as a result of increasing the pressure inside the cuff was observed to be 25% at 67 mm Hg. This test demonstrates the flexibility of the cuff and its response to nerve swelling post-implant. The stability of the contacts&#39; interface and recording quality were also examined with contacts&#39; impedance and signal-to-noise ratio metrics from a chronically implanted cuff (7.5 months), and observed to be 2.55 &#177; 0.25 k&#8486; and 5.10 &#177; 0.81 dB respectively.

This article provides a detailed description on the fabrication process of a high contact-density flat interface nerve electrode (FINE). This electrode is optimized for recording and stimulating neural activity selectively within peripheral nerves.

녹화 및 자극 애플리케이션을위한 높은 접촉 밀도의 제작, 평면 인터페이스 신경 전극

Many attempts have been made to manufacture multi-contact nerve cuff electrodes that are safe, robust and reliable for long term neuroprosthetic ...

Implantation of Chronic Silicon Probes and Recording of Hippocampal Place Cells in an Enriched Treadmill Apparatus

An important requisite for understanding brain function is the identification of behavior and cell activity correlates. Silicon probes are advanced electrodes for large-scale electrophysiological recording of neuronal activity, but the procedures for their chronic implantation are still underdeveloped. The activity of hippocampal place cells is known to correlate with an animal&#39;s position in the environment, but the underlying mechanisms are still unclear. To investigate place cells, here we describe a set of techniques which range from the fabrication of devices for chronic silicon probe implants to the monitoring of place field activity in a cue-enriched treadmill apparatus. A micro-drive and a hat are built by fitting and fastening together 3D-printed plastic parts. A silicon probe is mounted on the micro-drive, cleaned, and coated with dye. A first surgery is performed to fix the hat on the skull of a mouse. Small landmarks are fabricated and attached to the belt of a treadmill. The mouse is trained to run head-fixed on the treadmill. A second surgery is performed to implant the silicon probe in the hippocampus, following which broadband electrophysiological signals are recorded. Finally, the silicon probe is recovered and cleaned for reuse. The analysis of place cell activity in the treadmill reveals a diversity of place field mechanisms, outlining the benefit of the approach.

We describe the diverse steps to implant chronic silicon probes and to record place cells in mice that are running head-fixed on a cue-enriched treadmill apparatus.

만성 실리콘 프로브 및 녹음 풍부한 디딜 방 아 장치에 Hippocampal 장소 세포의 이식

An important requisite for understanding brain function is the identification of behavior and cell activity correlates. Silicon probes are advanced ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent development of thin, flexible ECoG electrodes has enabled conduction of stable recordings of large-scale cortical activity. We have developed a whole-cortical ECoG array for the common marmoset. The array continuously covers almost the entire lateral surface of cortical hemisphere, from the occipital pole to the temporal and frontal poles, and it captures whole-cortical neural activity in one shot. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain. Marmosets have two advantages regarding ECoG recordings, one being the homologous organization of anatomical structures in humans and macaques, including frontal, parietal, and temporal complexes. The other advantage is that the marmoset brain is lissencephalic and contains a large number of complexes, which are more difficult to access in macaques with ECoG, that are exposed to the brain surface.These features allow direct access to most cortical areas beneath the surface of the brain. This system provides an opportunity to investigate global cortical information processing with high resolutions at a sub-millisecond order in time and millimeter order in space.

We have developed a whole-cortical electrocorticographic array for the common marmoset that continuously covers almost the entire lateral surface of cortex, from the occipital pole to the temporal and frontal poles. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain.

일반적인 마모 셋에 전체 피 질 Electrocorticographic 배열의 만성 이식

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Wide-field Single-photon Optical Recording in Brain Slices Using Voltage-sensitive Dye

Wide-field single photon voltage-sensitive dye (VSD) imaging of brain slice preparations is a useful tool to assess the functional connectivity in neural circuits. Due to the fractional change in the light signal, it has been difficult to use this method as a quantitative assay. This article describes special optics and slice handling systems, which render this technique stable and reliable. The present article demonstrates the slice handling, staining, and recording of the VSD-stained hippocampal slices in detail. The system maintains physiological conditions for a long time, with good staining, and prevents mechanical movements of the slice during the recordings. Moreover, it enables staining of slices with a small amount of the dye. The optics achieve high numerical aperture at low magnification, which allows recording of the VSD signal at the maximum frame rate of 10 kHz, with 100 pixel x 100-pixel spatial resolution. Due to the high frame rate and spatial resolution, this technique allows application of the post-recording filters that provide sufficient signal-to-noise ratio to assess the changes in neural circuits.

We introduce a reproducible and stable optical recording method for brain slices using voltage-sensitive dye. The article describes voltage-sensitive dye staining and recording of optical signals using conventional hippocampal slice preparations.

전압에 민감한 염료를 사용하여 뇌 슬라이스의 광자 광자 광기록

Wide-field single photon voltage-sensitive dye (VSD) imaging of brain slice preparations is a useful tool to assess the functional connectivity in neural ...

Preparation of Peripheral Nerve Stimulation Electrodes for Chronic Implantation in Rats

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. Several recent studies have demonstrated the effectiveness of chronic VNS in enhancing central nervous system plasticity to improve motor rehabilitation, extinction learning, and sensory discrimination. Construction of chronically implantable devices for use in such studies is challenging due to rats&#8217; small size, and typical protocols require extensive training of personnel and time-consuming microfabrication methods. Alternatively, commercially available implantable cuff electrodes can be purchased at a significantly higher cost. In this protocol, we present a simple, low-cost method for construction of small, chronically implantable peripheral nerve cuff electrodes for use in rats. We validate the short and long-term reliability of our cuff electrodes by demonstrating that VNS in ketamine/xylazine anesthetized rats produces decreases in breathing rate consistent with activation of the Hering-Breuer reflex, both at the time of implantation and up to 10 weeks after device implantation. We further demonstrate the suitability of the cuff electrodes for use in chronic stimulation studies by pairing VNS with skilled lever press performance to induce motor cortical map plasticity.

Existing approaches for constructing chronically implantable peripheral nerve cuff electrodes for use in small rodents often require specialized equipment and/or highly trained personnel. In this protocol we demonstrate a simple, low-cost approach for fabricating chronically implantable cuff electrodes, and demonstrate their effectiveness for vagus nerve stimulation (VNS) in rats.

쥐에서 만성 이식을 위한 말초 신경 자극 전극의 준비

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. ...