Biology Reference
In-Depth Information
14.2.2
Devices
Fluorescence spectrometer (Fluorolog 3-2.2, HORIBA Jobin Yvon):
Double-grating emission monochromators, improving the system's stray light
rejection.
A PMT (950 V) with a high dynamic range (1-10
7
) that allows better
signal-to-noise ratio.
A swing-away mirror allows collection of sample luminescence at standard
right angle 90
to the excitation beam or front face at 22.5
.
XBO lamp (450 W).
FluorEssence version 3.0 software for data acquisition used FluorEssence-3.0
as a plug-in for OriginPro software.
Quartz cuvette 10 mm light path (Hellma, type no, 101-QS).
Inverted confocal laser scanning microscope—Zeiss LSM 780:
Axio Observer.Z1.
Laser lines: diode 440 (25 mW), argon 488 (25 mW) and 514 (25 mW), DPSS
561 (20 mW) for CFP-YFP, and GFP-mCherry derivatives.
Objectives: C-Apochromat 40
/1.2W DIC III, Plan-Apochromat 63
/1.4
/0.8 DIC II.
Dichroics (InVis and VisTwin gate main beam splitter (MBS)): MBS 440 and
458/514 for CFP-YFP and MBS 488/561 for GFP-mCherry FPs.
A motorized piezo XY stage (Merzh
¨
user).
Definite focus.
HBO lamp (100 W).
ZEN 2010 imaging software package.
Stage adapter (64-0327) and low-profile open bath chamber (RC-41LP)
(Warner Instruments).
Upright semiconfocal spinning disk microscope:
Spinning disk (CSUX1-M2N-E Yokogawa Electric Corporation).
Laser lines: diode 447 (50 mW) and 515 (50 mW) and DPSS 561
(50 mW).
Objectives: 60
Oil DIC II, and Plan-Apochromat 20
/1.00W LUMPlanFLN and 20
/0.95W XLUMPPlanFL
(Olympus).
OptoSplit II image splitter (CAIRN Research, UK) equipped with filterset
I (dichroic 510dcxr, CFP emitter 483/32 nm, and YFP emitter 542/27 nm) and
filterset II (dichroic 560dcxr, GFP emitter 537/26 nm, and mCherry emitter
609/54 nm).
NanoScanZ-100
m (Prior Scientific).
EMCCD iXon 897 with 95% Q.E and 35fps camera (Andor Technology).
HG lamp (100 W).
Andor iQ1.10.5 imaging software.
Three different kinds of black Teflon-coated inserts (Luigs & Neumann)
and 50 mm glass coverslip were glued with hot wax at the bottom of this
chamber.
m
