Operating Instructions for the BioPulverizer
The BioPulverizer reduces tissue frozen in liquid nitrogen or dry-ice to a course powder. The method is especially useful for tough tissue such as skin, cartilage, and cornea. Cryo-fracturing with the BioPulverizer is a preliminary step for cell disruption. Final disruption of the cells is accomplished using mechanical disrupters and/or aggressive extraction solutions (see homogenizer review at www.biospec.com).
1) The mortar is consits of two parts... an inner sleeve and an outer s.s. cup. The inner sleeve of the mortar nests in the outer cup. It is the hole in the inner sleeve that receives the tissue sample. Place the mortar and pestle of the BioPulverizer (BP) in an insulated container and thoroughly chill it with liquid nitrogen or, second best, crushed dry-ice. Stainless steel is relatively slow to cool down and later, slow to warm up. Slowly add liq N2 to the stainless steel parts until liq N2 no longer flashes off vapor and some liquid lingers on the stainless steel.
2) Add pre-frozen (-80 deg C or lower) tissue to the well of the mortar. Fresh tissue sticks to a cold BP on contact and cannot be inserted into the well.
3) Remove the BP from the insulated container and place it on the lab bench. There will be ample time to work with the chilled BP. Insert the pre-chilled pestle into the well of the mortar, patterned end down, and deliver one to three sharp blows to the pestle using the provided dead-blow hammer. Rotate the pestle slightly between blows.
4) Remove the pestle and the inner sleeve of the motar, thus providing easy access to the powdered sample.
IMPORTANT! Powdered tissue has a “dry” appearance and, with the exception of highly fatty tissue, can be poured into a test tube or plastic weighboat containing extraction solution. Pulverized tissue thaws quickly when exposed to room temperature, so work quickly or you will have a sticky mess.