Biology Reference
In-Depth Information
11. An acquisition system made up of an interface box (EMKA
Technologies, Cat.# itf16) connected to an acquisition card
(National Instruments, Cat.# PCI-6023E) inserted to a com-
puter (see Note 9).
12. A computer with an installed acquisition, registering and pro-
cessing software (iox version 1.8.9, EMKA Technologies)
(see Note 10).
1. Calibrating cylinders for gas analyzers zero: 100% N
2
, certified
(see Note 11).
2. Calibrating cylinders for gas analyzers span: 21% O
2
, 5% CO
2
,
and balance N
2
, certified.
3. Cylinders containing 15, 10, and 6% O
2
, respectively (each
with 5% CO
2
and balance N
2
).
4. Each gas cylinder should be equipped with a pressure regulator
and a manometer corresponding to the gas contained in the
cylinder.
5. Flexible PVC tubing nontoxic (conform to FDA standard) and
nonporous (see Note 12).
6. Nylon connector kit (Harvard Apparatus) (see Note 13).
7. Desiccant membrane air dryer (Perma Pure) (see Note 14).
2.2 Experimental
Gases
1. Sufficient number of adult mice (wild-type or transgenic) of a
comparable age and of determined sex to generate reliable and
reproducible mean group data (see Note 15).
2. Thermocouple monitoring thermometer and stainless steel
rectal probe for mice (see Note 16).
3. In case of experiments involving injection of exogenous Epo
(to wild type mice, typically), human recombinant Epo
(see Note 17).
2.3
Mice and Epo
3
Methods
1. Calibrate the air flow in to 700-800 ml/min by adjusting the
air supplying pump (see Note 18).
2. Calibrate the air flow out to 600-700 ml/min by adjusting the
subsampling pump (see Note 19).
3. Calibrate the CO
2
analyzer by using a 100% N
2
cylinder to cali-
brate the zero and a certified cylinder containing 5% CO
2
(with 21% O
2
and balance N
2
) to calibrate the span.
4. Calibrate the O
2
analyzer by using a 100% N
2
cylinder to cali-
brate the zero and a certified cylinder containing a percentage
3.1
Calibrations
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