Technical services

Experimental methods

• 免疫荧光 (IF) 通用

• 冷冻切片制备

• 动物行为学测定

  分析对象为自由活动动物在测试装置中的行为输出,随范式不同包括 Y 迷宫中的臂进入顺序与臂进入总次数、由连续三次进入构成的三连全臂进入(自发交替)、与进入总数对应的最大可能交替数,以及视频跟踪记录的移动距离等运动-选择指标。

• 单细胞转录组 scRNA-seq

• 反转录与 cDNA 合成

• 因果中介分析 (Mediation Analysis)

• 图像形态计量学

• 多剂量梯度给药

• 多细胞因子联检 (ELISA/Luminex)

• 天狼星红/Masson 纤维化染色

Experimental protocols

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

• 结肠大体肿瘤计数、卡尺直径测量与肿瘤负荷计算

  Version 1

  Tumor-burden endpoint for the AOM/DSS colitis-associated cancer model: at the day-100 endpoint mice are euthanized by cervical dislocation, the entire colon is excised into 15 ml cold sterile PBS, flushed with 5-10 ml cold PBS through a 21 G needle, opened longitudinally and rinsed again; macroscopic tumors are counted one by one, each tumor's diameter is measured with a digital caliper, and tumor load per mouse is the summed size of all its tumors. Photographed colons are fixed in 10% formalin solution (Sigma-Aldrich HT-501128) for paraffin sections and histological grading. Applies to AOM/DSS models (predominantly C57BL/6 background) for comparing tumor number, size distribution and intervention effects; the adopted record reports 100% tumor take in its in-house C57BL/6-background wild-type mice at day 100.

• 血样 ALT 活性测定（Teco A524 动力学法）

  Version 1

  Teco A524 liquid reagents quantify ALT activity from the decrease in NADH absorbance at 340 nm. The readout measures liver injury.

• DSS 结肠炎逐日 DAI 评分与粪便隐血检测

  Version 1

  Daily mouse colitis scoring combines body-weight loss, stool consistency and Hemoccult SENSA fecal occult blood testing. Each component ranges from 0 to 4 and the combined DAI ranges from 0 to 12.

• Hematoxylin and Eosin (H&E) staining for histopathology

  Version 14

  Hematoxylin and eosin (H&E) staining of fixed, paraffin-embedded tissue sections (4 μm). The workflow covers dewaxing, rehydration, hematoxylin nuclear staining, bluing, eosin counterstaining, dehydration and clearing, and coverslipping, yielding blue-purple nuclei and pink cytoplasm and connective tissue fibers for light-microscopic assessment of cellular and tissue morphology.

• In Vivo Checkpoint Antibody Administration in Mouse Tumor Models

  Version 3

  In vivo checkpoint antibody dosing SOP for mouse subcutaneous syngeneic tumors: cell culture and Day-0 implantation (MC38/B16-F10 in C57BL/6 females, CT26/4T1 in BALB/c females, 6-8 weeks); caliper measurement every 2-3 days with volume V=(LxW^2)/2 and body weight; volume-based randomization; i.p. anti-PD-1 (RMP1-14, 200 ug/mouse, days 0/3/6/9), anti-CTLA-4 (200 ug/mouse, days 0/3/6) or combination with isotype controls; readouts: TGI (above 60% biologically significant), body weight, humane endpoints (excessive tumor size, ulceration, marked weight loss), terminal analysis (weighing, 10% formalin fixation for IHC, dissociation flow cytometry, serum cytokines).

• 大鼠尾静脉血糖测定

  Version 9

  Measure rat tail-vein blood glucose using Roche Accutrend Plus test strips and meter.

• 小鼠胰岛素耐量试验（ITT）

  Version 7

  This protocol adopts the protocols.io Insulin Tolerance Test in Mouse v1. Mice are fasted for 4–6 hours, weighed, and given insulin intraperitoneally at 1–1.2 U/kg. Blood glucose is measured before and at 15, 30, 60, 90 and 120 minutes after injection. The measurement is a glucose assay (OBI:2100020); insulin administration, dose and time points belong to this concrete protocol and are not presented as an exact OBI insulin-tolerance-test term.

• 小鼠腹腔注射葡萄糖耐量检测（IPGTT）

  Version 8

  Intraperitoneal glucose tolerance test (IPGTT) in mice: single-house for one week, weigh, fast for 6 hours, measure baseline glucose, inject 2 g/kg body weight of 20% D-glucose intraperitoneally, then collect blood at 15 to 120 minutes after injection. Used to assess glucose clearance in obesity and glucose-metabolism models.

• 小鼠口服葡萄糖耐量试验（OGTT）

  Version 14

  This protocol adopts the protocols.io Oral Glucose Tolerance Test in Mouse v1. Mice are fasted for 4–6 hours, weighed, and given oral glucose at 1 g/kg (0.5 g/kg after at least 4 weeks of high-fat diet). Blood glucose is measured before and at 15, 30, 60, 90 and 120 minutes after the load, with tail-vein plasma collected at selected time points for insulin measurement. The record describes the assay procedure, not executed disease-model results.

• 天狼星红（Picrosirius red）胶原染色

  Version 7

  This protocol adopts the protocols.io Picrosirius red staining for histology v1. Formalin-fixed paraffin-embedded sections are dewaxed and rehydrated, optionally pre-treated with phosphotungstic acid and counterstained with Weigert haematoxylin, then stained with 0.1% Sirius Red F3BA in saturated picric acid, differentiated with acetic acid, dehydrated, cleared and mounted. Collagen appears red in bright field and birefringent under polarised light. The record describes staining and microscopic observation, not a complete fibrosis area or stage quantification.