Name: Acquisition and Pre-Processing of Nasal Mucosal Samples from Rats with Allergic Rhinitis
Uploaded: 2023-09-22T14:50:00
Description: Watch this Scientific Journal Video about Acquisition and Pre-Processing of Nasal Mucosal Samples from Rats with Allergic Rhinitis at JoVE.com

acquisition and pre-processing of nasal mucosal samples from rats with allergic rhinitis

アレルギー性鼻炎ラットモデルにおける鼻組織の多色免疫蛍光法

Allergic rhinitis (AR) is a chronic, non-infectious inflammatory disease of the nasal mucosa primarily mediated by specific immunoglobulin E (IgE)1,2. It is characterized by symptoms such as sneezing, a runny nose, nasal congestion, and nasal itching. With industrialization and urbanization, the prevalence of AR is gradually increasing, affecting approximately 10%-20% of the world&#39;s population1. The immunofluorescence (IF) technique is a fluorescent staining method that utilizes an antibody-antigen binding reaction. It can be employed to detect and quantify the distribution and expression levels of specific proteins in biological tissues or cells. In AR research, IF can simultaneously detect multiple targets, including AR-related cytokines, inflammatory cells, receptors, and more, facilitating the exploration of AR pathogenesis and the effects of drugs3,4,5,6.
The multicolor immunofluorescence (mIF) staining process closely resembles traditional IF, with the addition of an antibody elution step during each round of staining. This modification enables the simultaneous detection of multiple biomarkers on the same tissue section through sequential single-labeling and multiple rounds of re-staining. mIF is based on tyramide signal amplification (TSA), allowing for repeated cycles of TSA fluorescent staining and the use of microwave heating to remove antibodies while retaining fluorescent signals7,8. In comparison to conventional IF, mIF offers several advantages: (1) it can detect weakly expressed antigens that are challenging to identify with conventional IF9,10; (2) it provides high-quality staining with an improved signal-to-noise ratio; (3) it allows for the quantification of tissue-specific structures and regions of interest11; (4) multiplexing multiple pathways efficiently utilizes tissues and conserves limited pathological resources12; (5) multi-parameter analysis through mIF offers deeper insights into tissues, uncovering hidden biological information13.
Overall, mIF allows the observation of different antigen expressions and distributions within the same sample, facilitating the study of target proteins. In the future, researchers seeking to understand the expression and distribution of multiple target proteins will find this technique a valuable choice. This study demonstrates the application of mIF for staining nasal mucosa samples from rats with AR and evaluate the establishment of a rat model of AR.

著者スポットライト:多色免疫蛍光によるアレルギー性鼻炎研究の進歩

多色免疫測定法 による アレルギー性鼻炎ラットの鼻粘膜組織切片における免疫蛍光標識

Our main research is based on a study of allergic rhinitis. Our research intends to analyze what proteins are associated with allergic rhinitis. Multicolor immunofluorescence is currently used to advance research in our field.In future studies, we will focus on the possible mechanisms of TCM in the treatment of allergic rhinitis.

Watch this Scientific Journal Video about Acquisition and Pre-Processing of Nasal Mucosal Samples from Rats with Allergic Rhinitis at JoVE.com

Acquisition and Pre-Processing of Nasal Mucosal Samples from Rats with Allergic Rhinitis  

アレルギー性鼻炎ラットからの鼻粘膜サンプルの取得と前処理

まず、メスで、安楽死させたラットの口の2つの角に沿って正確に切開し、筋肉組織を露出させます。組織はさみを使用して、頬骨の関節と頭蓋骨の両側の下顎骨を切断します。次に、下顎骨を取り除きます。ティッシュハサミを使用して、鼻腔と上顎骨の間の骨の接続を切断します。次に、止血剤を使用して上顎骨の皮膚を分離し、鼻腔を露出させます。眼窩下孔で鼻腔と両側の眼窩骨との接続を切断し、鼻腔を切除します。抽出した鼻腔を4%パラホルムアルデヒド溶液に入れて固定します。組織を脱灰するには、固定された組織を蒸留水で洗浄します。室温で組織の20〜30倍の体積の脱灰溶液に組織を入れます。脱灰が完了したら、組織を脱水ボックスに入れます。脱水ボックスを脱水機に移し、異なるインキュベーション時間でアルコール濃度を上げて組織を脱水します。次に、組織を無水エタノールに30分間2サイクル入れます。これに続いて、組織をキシレンに5〜10分間移して繰り返し、次に組織をワックスに1時間浸します。ワックスブロックを埋め込みカセットに入れます。ワックスブロックが固まったら取り除き、ワックスブロックをトリミングします。トリミングしたワックスブロックをパラフィンミクロトームに挿入し、厚さ3マイクロメートルのスライスにカットします。対照ラットのヘマトキシリンとエオシン染色は、よく整理され、繊毛を示しました。鼻中隔粘膜は損傷を受け、好中球浸潤が著しい疾患群で剥離した。

Research

JoVE Journal

Medicine

Acquisition and Pre-Processing of Nasal Mucosal Samples from Rats with Allergic Rhinitis

Immunofluorescent Labeling in Nasal Mucosa Tissue Sections of Allergic Rhinitis Rats via Multicolor Immunoassay

Allergic rhinitis (AR) is a chronic, non-infectious inflammatory disease of the nasal mucosa, primarily mediated by specific immunoglobulin E (IgE), ...