Hello Archana Tripathi

Fresh tumor tissue should be used for any type of application such as for tissue transplantation and for nucleic acid isolation. For high quality RNA isolation, it is imperative that the tissue be flash frozen within 10-20 min after removal from the body to prevent degradation. For tissue transplantation, time is less critical, but the tissue should remain on ice and in medium until transplantation.

The materials that you will require are gloves, disposable bibs, sterile gauze, swabs, long, straight trimming blades, sterile scalpels, tissue culture dishes, sterile forceps, 50ml conical tubes, RNase/DNase-free cryovials, bleach, 70% ethanol, RNAlater (RNA Stabilization Reagent) and Dewar of liquid nitrogen.

You will have to coordinate tissue collection. The tissue collection staff checks the surgery schedule daily to identify the participants. The tissue collection staff also reviews and notes the patient history and the clinical diagnosis. A tissue collection coordinator alerts the surgeon, pathologist, investigator and tissue repository staff by e-mail 24 hr. prior to surgery.

Informed consent must be obtained from the patient prior to tissue collection. Prior to the surgical procedure, the repository technician communicates with the operating room staff that a specimen for research has been planned. The operating room staff notifies the tissue repository technician 10 min prior to when the tissue is removed from the patient. This will ensure that the repository technician can get to the operating room before the tissue is removed from the patient in order to minimize the amount of time prior to processing.

When the breast tissue is removed, the technician notes the time of the day, and places the tissue into a container on ice for immediate transport to the pathology laboratory. The specimen is potentially infectious and should be handled with blood-borne pathogen considerations. The pathologist verifies the patient identification and removes surplus tumor and adjacent uninvolved (normal) tissue for research purpose. The tissue type (cancerous or adjacent uninvolved), location within the breast from where the tissue was removed, and procurement time are noted on a specimen worksheet.

From each tissue about 30 mg pieces are placed into separate cryovials, and immediately flash frozen in liquid nitrogen. The frozen tissues are stored in vapor phase liquid nitrogen. You could also make additional 20 mg pieces from the tissue and place into separate cryovials containing 400 μl of RNAlater for RNA analysis. These tubes are stored at 4°C overnight. You could remove the RNAlater as per manufacturer’s directions and store the specimens at −80°C.

If you are left with extra tumor specimen, you could place the tissue into separate 50 mL conical tubes containing the growth media for immediate tissue implantation or disassociation to organoids if you are planning for such experiments.

It is important that you need to enter patient identification, specimen collection information, as well as storage information in a biospecimen tracking database.

Good luck!

We had experience of working with oncosurgeon. The most important thing we had to deal with was the timing. Sometimes, our fellow had to wait for a tissue more than 12hrs. We don't have -80 fridges at tissue collection site and don't have that much facilities. For tissue sample preservation, RNALater is a blessing for us. Part of the tumor tissues was kept for pathological tests and the rest was given to us for molecular analysis. Tissue sample was stored in RNALater containing tube immediately after collection, transported to lab in an icebox, and stored at -20C.

Collecting tumor samples from breast cancer patients requires careful adherence to protocols and ethical considerations. Here is a detailed protocol for tumor sample collection from breast cancer patients, along with necessary precautions to be taken:

Remember to consult and collaborate with experienced researchers, pathologists, and institutional review boards to align your protocols with local regulations and best practices specific to your research setting.

It is crucial to prioritize patient safety, privacy, and ethical considerations throughout the entire process of tumor sample collection.