Task 2 21-Oct-2020
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 maximal intensity
of fluorescence of your fluorophore?
Please, select the nearest laser which is available in SpectraViewer
application in Light Sources.
3. Which of installed long-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 to select suitable emission
filter and design band-pass filter use SpectraViewer application
http://www.invitrogen.com/site/us/en/home/support/Research-Tools/Fluorescence-
SpectraViewer.html
Instructions: Chose your flurophore. Insert a suitable available laser in Light
Sources by selecting Custom. Enter laser wavelength value. Show a suitable filter
in Emission Filters by selecting Custom. Enter filter wavelength value. In Badwidth
enter LP for longpass filter or filter range in nanometers for bandpass filter.
Fluorophore A can be found in the table next to your name.
Please send me your short answers via email within 48 hours.
A correct answer = 0.5 point
A
1 Dikunová Alžbeta FITC (Fluorescein)
2 Dzurov Matej ROX (carboxy X rhodamine)
3 Faturová Jana Acridine Orange
4 Gašparik Norbert GFP (emerald GFP)
5 Hesko Ondrej Lucifer Yellow
6 Jahodová Kateřina Ethidium Homodimer
7 Kameniarová Michaela Pacific Green
8 Konečná Kateřina JOE
9 Korytářová Anna Alexa Fluor 488
10 Kozeleková Aneta Cy3
11 Kubinyiová Lenka Oregon Green 514
12 Kůřilová Eliška Cyanine5
13 Lysáková Klára SYBR Green
14 Mikšátková Barbora Red Fluorescent Protein (RFP)
15 Nováková Barbora TOTO-3
16
Prabhullachandran
Unnikannan
Alexa Fluor 594
17 Procházková Markéta BOBO-1
18 Šimek Jan Sypro Orange
19 Tužinčin Dávid TOTO-1