Regarding the WLL, keep in mind it is a 80 MHz pulsed laser. Therefore, you can do time domain FLIM or gating with excitation at any wavelength from 440 to 790 nm.
With a detector sensitive out to 850 nm, this is also a way to add functionality for new far red probes.
Cheers-
Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
NYU Langone Health, 540 First Avenue, office MSB 0657A (elevator to MSB 6 or Smilow 4)
main lab Smilow C-17, New York, NY 10016
Office: 646-501-0567 Cell (DO NOT TEXT): 914-309-3270 Michael.Cammer@med.nyu.edu
http://nyulmc.org/micros http://microscopynotes.com/
Scheduling the time you want is far more reliable by phone call. Why not provide your phone number?
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Original Message:
Sent: 3/18/2024 8:35:00 AM
From: Anthony Stender
Subject: RE: Are White Light Lasers worth the money?
Thank you, Mahmud, for that perspective. It was quite helpful. For what it's worth, if we purchase a WLL, it would be part of a Leica system.
My primary concerns were the cost of a service contract and laser power. I have previously heard that the tradeoff for a WLL is power, so I'm glad you addressed that. I'm continuing to look into the cost of a service contract, since our core needs to keep that in mind for any new purchase. It's also helpful to hear about your experience with laser repair. That was something I didn't think about, but it is definitely something I need to keep in mind.
At this point, I am simply doing some fact-finding. It is unclear if there will be enough support for a WLL, at least in the short-term.
Anthony
Original Message:
Sent: 3/11/2024 1:14:00 AM
From: Mahmud Siddiqi
Subject: RE: Are White Light Lasers worth the money?
I have two Leica confocals with WLLs (NKT Photonics), both acquired with the systems through Leica. In my experience, there are only a few 'cons' --
Initial costs: A WLL can add $100K - $200K to the cost of a full system, when compared to a system with fixed-wavelength diode lasers. However, this is not so different from adding a laser for multiphoton fluorescence. Both lasers are bundled into the confocal system service contract, so I am unable to give you numbers on a service agreement for the laser in isolation.
Repair turnaround times: In the rare instance (twice in ~9 years) that the WLL needs factory repairs, the turnaround time can be long (weeks - months). In my case, this was not overly painful, as a loaner WLL was available.
Somewhat lower output power at selected wavelengths: This is less of an issue with newer versions. While diodes can provide more mW at their specific wavelengths, this is rarely an issue for biological specimens which can withstand only a fraction of the maximum available excitation laser power in any case. If higher powers are required, e.g. for photobleaching, applying e.g. 512, 514, 516 nm simultaneously delivers more than enough energy.
Tuning range: Again, less of an issue with newer versions. On my older system I have 405nm and 445nm diodes in addition to the WLL (470-670nm); on the newer one only the 405 nm diode is required (WLL 440-830nm).
A sort of 'global con' is that the rest of the confocal system has to be able to take advantage of the various capabilities and characteristics (tunability, pulse synchronization, etc.) of the WLL to make the purchase really worth the price, especially if you will be adding only the laser. What kind of confocal would you be retrofitting?
An argument can be made that having the ability to output 517nm instead of 514nm is a luxury (although I did have to do exactly that for a recent imaging experiment). However, if your users bring you uncharacterized fluors, or unusual fluor combinations, it's very hard to go back to fixed laser lines after using a WLL.
If there's a specific drawback of concern, or any specific questions, I'll be happy to elaborate as best I can...
Mahmud
Mahmud Siddiqi, PhD
Imaging Facility Manager
Carnegie Institution / Embryology