Notes for ISH services

1. Please go to the form download page to download and fill out the application form.
2. If user wants to collect sample by themselves, please provide more than 5 tissues/ sample. Please use a sharp and clean razor or scalpel to sample freshly and place it in freshly prepared PFA fixative (4% paraformaldehyde + 0.1%Tween 20 + 0.1% TritonX-100 + 0.25% glutaldehyde/ 1XPBS), apply a vacuum at 4°C (680~720 mmHg) for several hours (depending on the sample). After the sample is taken out, use 1xPBS Wash once, place on ice and replace with 30%, 50% and finally 70% alcohol every 30 minutes, then send to our laboratory at low temperature. If sampling is taken by ISH CORE, please clearly indicate the location of the sampling and whether there are any special sampling period requirements for the materials.

If the special sample* failed in sectioning twice, the case must be closed first, and then we can discuss ways to improve it.

*Special samples are considered to meet any of the following conditions: the length, width, and height of the sample are less than 1mm, the designated observation area is less than 0.5mm, and special angle of tissue sectioning. 

3. The length of the hybridization probe is recommended to be 150~250bp. Please clone it in pGEM T-easy vector. Inform us the     target gene whether the forward direction is from T7 or SP6 promoter, and whether the probe contains restriction enzyme   cleavage sites.

4. If you prepare your own Plasmid DNA, the total amount should be > 20ug, DNA conc.＞700ng/ul (Quality O.D. Ratio A260/A280= 1.8-2.0).

5. If samples or plasmid DNA are shipping over long distances, please package them carefully to avoid crushing.  

6. The probe design must be gene specific and have high gene expression level (please attach qPCR and other experimental results for reference); our CORE will  test four probe hybridization temperatures, two probe concentrations, and repeated it three times. If non-specific background cannot be avoid, the probe is deemed to be inapplicable and the case needs to be closed and pay some processing fee.   

7. ISH core provides quality control photos taken under a microscope (no extra charge) to confirm that the experimental result is correct. The objective magnification of the QC photos is determined by the sample size, and not include photos of the entire tissue. 

8. Quality control photo files are save in tif files and sent by e-mail. All slides used for the experiment will be returned to the user. Our laboratory only provides experimental technical services and is not responsible for taking the microscopic photography of the entire tissue and the interpretation of gene expression results. 

Submitting documents is deemed to be the establishment of a contract. If the contract is terminated midway, you will also have to pay some processing costs.