1 To begin, transfer the tumor tissue2 immersed in 25 milliliters of hibernate A 3 into a sterilized glass petri dish 4 under a laminar flow cabinet. 5 Using a microscope, carefully eliminate 6 all the necrotic tissue. 7 Then use a scalpel 8 and tissue forceps to dissect the blood vessels out.
9 Cut the tumor tissue into 10 one to two cubic centimeter pieces. 11 Transfer them into a plastic petri dish 12 containing three milliliters of H-GPSA medium, 13 and place the dish on ice. 14 To manually process the tumor, transfer the tumor piece 15 to a fresh glass petri dish.
16 Using two scalpels, dissect the segment 17 into 0.5 milliliter sections under a microscope. 18 To mechanically process the tumor, first, 19 position the blade properly. 20 Then adjust the slice thickness to 0.45 to 0.50 millimeters, 21 and fix the table release knob to start mode.
22 Cut the agaro block into a two centimeter-long cylinder 23 and glue it onto the circular plastic dish of the chopper. 24 Using a scalpel, create a pit in the agaro cylinder 25 and place the tumor tissue in the pit. 26 Then position the plastic disc onto the mounting disc 27 of the cutting table, and press the reset button 28 to start the chopping process.
29 After the first round, rotate the mounting disc 30 by 90 degrees, and repeat again 31 to cut the tissue into rectangular pieces. 32 Remove the plastic disc. 33 And then using a tissue spatula, 34 rotate only the tumor tissue by 90 degrees.
35 Return the plastic disc to the cutting table 36 and press reset to continue cutting. 37 Then using a five milliliter pipette, 38 transfer the processed material, 39 along with the medium, to the petri dish. 40 Place the dish immediately on ice.
