Technical services

Experimental methods

• qPCR/WB 免疫检查点验证

• RNA测序

• RT-PCR

• RT-qPCR 通用

  测定对象为反转录产物中目标序列的实时扩增量，原始观测为循环阈值（Cq），需经扩增效率校正后转换为可比较的目标量。

• Sanger sequencing

• STZ 注射诱导糖尿病

• TILs 多色免疫组化 (mIHC)

• transcription profiling assay

• triglyceride assay

• TTC 梗死体积染色

Experimental protocols

Protocols apply a method to a defined measurement, with samples, procedures and quality controls. Available services are linked separately.

• 脑切片 TTC 染色与梗死面积测定

  Version 8

  Fresh brains are cut to 2-mm coronal sections, stained per well with 2% TTC in 0.1M PBS at 37°C for 30 min protected from light, stored in fixative, imaged on a stereomicroscope, and infarct percentage computed ipsilateral-versus-contralateral.

• 大鼠 Morris 水迷宫空间定位与再学习测定

  Version 3

  A rat water-maze procedure using an automated Atlantis platform and distal cues, with a new platform location each day. Preserve the separate platform-found and platform-not-found paths, and record escape latency and swim trajectories across the successive trials.

• 脑切片酪氨酸羟化酶（TH）免疫荧光染色

  Version 5

  Brain sections are washed with 4% PFA in PBS, blocked with 10% donkey serum/0.3% Triton-X, incubated with chicken anti-TH ab76442 (1:200) and a chicken-IgG fluorescent secondary antibody (1:1000), mounted in Mowiol and imaged by fluorescence microscopy to localize TH-positive cell bodies and fibers.

• 脑切片酪氨酸羟化酶（TH）免疫荧光染色

  Version 4

  Brain sections are washed with 4% PFA in PBS, blocked with 10% donkey serum/0.3% Triton-X, incubated with chicken anti-TH ab76442 (1:200) and a chicken-IgG fluorescent secondary antibody (1:1000), mounted in Mowiol and imaged by fluorescence microscopy to localize TH-positive cell bodies and fibers.

• 脑组织 RNA 第一链 cDNA 合成

  Version 1

  Synthesize first-strand cDNA from purified brain total RNA using ProtoScript II reverse transcriptase.

• 脑组织总 RNA 提取与柱上 DNA 去除

  Version 1

  Extract total RNA from brain tissue using TRIzol phase separation, a silica spin column and on-column DNase treatment.

• 大鼠脑组织 Sirt1 转录本定量（cDNA qPCR）

  Version 1

  Quantify Sirt1 relative to GAPDH in cDNA prepared from rat brain RNA using separate SYBR Green qPCR reactions.

• Y 迷宫自发交替检测

  Version 3

  This protocol records 8 min of free exploration in a symmetrical Y-maze to measure spontaneous alternation as an index of working memory: mice acclimate in the behavior room for at least 01:00:00, are placed in the center of a maze cleaned with 70% ethanol and dried, and arm entries are video-tracked with EthoVision; percent alternation equals alternations over max entries, with a correlation check between entries or distance traveled and alternation to detect activity confounds.

• FFPE 脑切片 6E10 碱性磷酸酶免疫组织化学检测 Aβ/APP

  Version 5

  This protocol uses the mouse monoclonal antibody 6E10 for alkaline-phosphatase immunohistochemistry on 2–3 µm FFPE brain sections, including dewaxing and rehydration, heat antigen retrieval in citrate buffer, formic acid treatment, hydrogen peroxide and normal goat serum blocking, overnight primary antibody incubation, alkaline-phosphatase secondary antibody incubation, FastBlue development and hematoxylin counterstaining. 6E10 recognizes beta-amyloid epitopes including APP, so the signal is not equivalent to insoluble plaque alone and does not give absolute Aβ40/Aβ42 concentrations. The validated source material is FFPE human brain; application to mouse brain expressing human or humanized Aβ requires endogenous-IgG and negative controls, and that boundary must remain in interpretation.

• 小鼠海马切片PSD95免疫荧光检测

  Version 1

  PSD95 immunofluorescence in prepared mouse hippocampal sections with DAPI nuclear staining.