Bio-Rad PDS-1000 / He™ and Hepta™ Systems - Manual (Page 7)

The helium pressure regulator (supplied) has a CGA 580, female fitting (standard in the
United States) for attachment to the user-supplied helium tank. An adaptor to this fitting may
be required outside the United States. Contact your local Bio-Rad office for information on
the helium pressure regulator adaptor requirements in your location.
A user-supplied, 1 inch adjustable wrench is required for attachment of the regulator to
a helium tank having a capacity of 55 cubic feet or greater.
Vacuum Source
The main unit of the PDS-1000/He system must be connected to a vacuum source capa-
ble of evacuating the bombardment chamber to a minimum of 5 inches of mercury for oper-
ation. This minimum vacuum requirement is part of the instrument safety system (the chamber
door must be sealed for helium pressure to be delivered into the main unit). Tubing is supplied
to connect the PDS-1000/He system to vacuum source.
For maximum evacuation capacity, we recommend connecting the PDS-1000/He device
to an oil-filled, rotary vane vacuum pump, either single or dual stage, with an pumping speed
of 90–150 liters/minute (3–5 cubic feet/minute). This pumping rate minimizes the time target
cells are exposed to vacuum. Oil for the vacuum pump must be supplied by the user. An
exhaust mist eliminator on the vacuum pump is also recommended.
The level of the vacuum required in the bombardment chamber depends on the biologi-
cal system being targeted for transformation. Higher vacuum reduces drag forces on micro-
carriers during helium-driven acceleration. During the brief bombardment process (less than
1 minute), typical protocols require a vacuum level within the bombardment chamber between
15–29 inches of mercury. Some cells, tissues, and intact plant cells require a high vacuum
(up to 28 inches of mercury) for efficient transformation.
House vacuum may be sufficient for bombardment of certain cell types (mammalian
cells), but house vacuum pumping rates can fluctuate and vary greatly in overall evacuation
capacity (typically 20 inches of mercury, maximum).
Consumables
The 500 Optimization Kit (catalog number 165-2278) provides the consumables needed
for 500 bombardments. It is recommended for users who have yet to determine the optimal
conditions for the bombardment of the biological system of interest. The kit contains 0.25 g
each of 0.6 µ, 1.0 µ, and 1.6 µ gold microcarriers, 100 each of the nine different rupture disks
(ranging from 450 psi to 2,200 psi), 500 macrocarriers, and 500 stopping screens. After opti-
mal conditions are determined, Standard Pressure Kits are available (see Section 7 or the cur-
rent Bio-Rad catalog for a complete listing).
Additional Laboratory Supplies and Equipment
Vortex mixer is needed for microcarrier preparation.
Common laboratory supplies, such as 95% ethanol, pipettes, etc. are required, as cited in
standard protocols.
Additional Items Available from Bio-Rad
The Yeast Optimization Kit (catalog number 170-3100) allows first-time users to become
familiar with the Biolistic instrument. It is also helpful for experienced users wishing to
periodically standardize their bombardment conditions. This kit provides all of the biological
material needed to transform yeast. Yeast provides a system that is quickly and easily assayed.
The kit demonstrates the effect of varying the DNA concentration, rupture disk pressure, and
the target distance (from the stopping screen), and includes
Saccharomyces cerevisiae
strain
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