BioPulverizer Instructions

The BioPulverizer reduces tissue frozen to liquid nitrogen or dry-ice temperatures 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 https://biospec.com/resources/cell-disrupters-a-review).

Operating

1. Place the assembled BioPulverizer in a small plastic container and thoroughly chill it with liquid nitrogen or, second best, crushed dry-ice. Stainless steel is slow to cool, but once cold, maintains the low temperature well. Slowly add liquid nitrogen to the stainless steel parts until nitrogen gas no longer flashes off and some liquid nitrogen lingers on the stainless steel.

1. Add appropriate sized pre-frozen (-80 °C or lower) tissue to the well of the mortar.  Fresh tissue sticks to the cold BioPulverizer on contact and cannot be inserted into the well.

1. Remove the BioPulverizer from the plastic cooling container and place it on the lab bench.  There will be ample time to work with the chilled BioPulverizer.  Insert the chilled pestle into the well of the mortar and deliver one to three sharp blows to the pestle using the dead-blow mallet.  Rotate the pestle slightly between blows for best results.

1. Powdered tissue has a “dry” appearance and can be poured into a test tube or plastic weigh boat containing extraction solution.  If the tissue was high in fat, the pulverized tissue may be in the form of a pellet or tablet. Pulverized tissue thaws quickly so work quickly or you will have a sticky mess.

Cleaning

Despite its name, “stainless” steel can be attacked by biomaterials.  Clean the BioPulverizer promptly.  After thawing the metal parts in warm water, wash the BioPulverizer using detergent.  For those using PCR techniques, a one minute exposure to a 1 to 10 dilution of household bleach (Clorox, etc.) destroys all residual nucleic acids – reference available.  Rinse thoroughly and dry.

IMPORTANT! Powdered tissue has a “dry” appearance and, with the exception of highly fatty tissue, can be poured into a test tube or plastic weigh boat containing extraction solution.  Pulverized tissue thaws quickly when exposed to room temperature and humidity, so work quickly to avoid a sticky mess.

Use caution when handling liquid nitrogen.  Always wear safety glasses and gloves.