4. Dulbecco’s phosphate buffered saline (DPBS): 8 g/L sodium
chloride, 200 mg/L potassium chloride, 1.44 g/L sodium
phosphate dibasic, 240 mg/L monopotassium phosphate,
pH 7.4.
5. 1 mL single-use syringe.
6. Ventilated 25 cm
2 flask.
7. 50 mL tube.
8. Petroff-Hausser chamber.
9. 24-well black plate.
10. 96-well black plate.
11. Adult mice, preferably white, with no age restriction or sex.
12. D-Luciferin: 30 mg/mL solution in DPBS. Aliquot 600 μL of
D-luciferin solution per tube.
13. Luminometer.
14. IVIS Spectrum imaging system. Combines 2D optical and 3D
optical tomography in one platform (Fig. 1).
15. XGI-8 Gaseous Anesthesia System (Fig. 2).
3 Methods
3.1 Leptospire
Culture
Leptospira strains are grown in liquid EMJH medium at 30
C with
no agitation. The generation time is 18–20 h for pathogenic
L. interrogans and 4–7 h for saprophytic L. biflexa strains.
Custom lens,
24-position
filter wheel
assembly,
laser light
alignment
Gas anesthesia
manifold
Compartment
for CCD camera
refrigeration
and
camera controller
Imaging
chamber
(heat pad)
CCD
camera
Scanning
laser
CCD dimension
image focus plane
lens aperture
object focus plane
field of view
filters
XGI-8 Gas
anesthesia system
Monitor
Acquisition
computer
Induction
chamber
Fig. 1 Schematic picture of the IVIS Spectrum system. Representation of the in vivo imaging system combining
2D optical and 3D tomography in one platform. Mice are anesthetized before imaging in the induction chamber
and during imaging by using the XGI-8 Gas Anesthesia System, designed to work with the IVIS Spectrum
[11]. On the right is a schematic representation of the major components for bioluminescence acquisition and
analysis using the in vivo imaging software [3]
In Vivo Imaging of Bioluminescent Leptospires
151