Task 2 6.11.2012
Selection of excitation lasers and emission filters
You came for a research stay into a renowned foreign laboratory. A principal
investigator showed you all instrumentation. When showing fluorescence scanner
for detection of electrophoretic gels, he asked you to write down equipment that you
would need for your experiments. As you will use fluorophore A, you need to find
out whether it is possible to excite and selectively detect fluorescence of your
fluorophore.
Excitation wavelengths of lasers installed are: 473 nm, 532 nm, 635 nm.
Installed long-pass emission filters are 510 LP, 575 LP, 665 LP.
1. Which laser would you chose for excitation of your fluorophore?
2. What laser would you order to obtain maximal excitation and also maximal
intensity of fluorescence of your fluorophore?
Please, select the nearest laser which is available in SpectraViewer
application.
3. Which of installed band-pass filters would be suitable for the detection of your
fluorophore?
4. What band-pass filter would you order for optimal detection of your
fluorophore? Please, write mean wavelength, dash band width (e.g. 530/20).
To see spectral characteristics of your fluorophore and design of band-pass filter,
use SpectraViewer application
http://www.invitrogen.com/site/us/en/home/support/Research-Tools/Fluorescence-
SpectraViewer.html
Note: For launching of the application is necessary to have Java installed in your
web browser.
Click on Toggle Excitation Display” for actualization of your spectra and filter
display.
Fluorophore A can be found in the table next to your name.
Please send me your short answers via email within 48 hours.
Correct answer = 0.5 point
A
1 Bencúrová, Petra Fluorescein (antibody conjugate)
2 Dabravolski, Siarhei 5 ROX (carboxy X rhodamine)
3 Dubec, Vít Acridine Orange
4 Dudová, Zdenka GFP
5 Dvořák, Jan Lucifer Yellow
6 Fabišik, Matej 5 FAM (carboxyfluorescein)
7 Fedorko, Jan 6 JOE
8 Fialová, Martina AlexaFluor 488 (oligo conjugate)
9 Holek, Michal Cy3 dye (antibody conjugate)
10 Kočka, Martin Oregon Green 514 (antibody conjugate)
11 Míka, Matěj Cy5 dye (antibody conjugate)
12
Obacz, Joanna
Agnieszka
Propidium iodide
13 Partyka, Jan YOYO-1
14 Přikrylová, Terézia SYBR Green-1
15 Rájecký, Michal YFP
16 Reichman, Pavel SYBR Gold
17 Sochorová, Jana FITC (antibody conjugate)
18 Škubník, Karel SYPRO Ruby
19 Tylichová, Zuzana RFP