Cameras for ultra low light levels
Posted in Hardware
Many image sensors are built backwards, just like the retina. The actual photon detectors are buried behind a mesh of circuitry. Back-illuminated sensors, where the detectors are put in…
Tag: imaging
More 2p Cross Sections
Posted in Tips
Here are some 2p cross sections for Alexa dyes from Invitrogen. Dye manufacturers are getting better about sharing 2p cross sections, but they still rely on out side people to do the measurements. In this case, Warren Zipfel from Cornell.
Speaking of…
Two-Photon Cross Sections
Posted in Tips
It’s difficult to track down 2p absorption cross sections for dyes and proteins. Doing it right requires (ideally)…
PMTs for 2-photon imaging
Posted in Hardware
On the topic of custom two-photon imaging rigs, what kind of PMTs do you want to go with? Naturally, you’ll order yours to be hand selected for high quantum efficiency and…
Optomechanics for a custom 2p uncaging rig
Posted in Hardware
Post by Darcy Peterka (Columbia University/Yuste lab):
We do a bunch of multiple imaging/uncaging experiments with 2+ lasers, and got tired of constantly taking apart the camera/laser ports to switch lenses and dichroics. This…
Building a 2-photon microscope
Posted in Hardware
Less than a year ago, the patent on 2-photon microscopy ran expired (Nov 14, 2009 pdf). So now anyone can sell a kit or system for two photon microscopy without having to pay a licensing fee.
If you don’t want to start from scratch, there are a lot of options.
Prairie Technologies
Lasers for multiphoton microscopy
Posted in Hardware
Coherent just sold its 1000th Chameleon system. That’s roughly how many of the similarly-priced Lamborghini Countach LP500s were sold. That was in the 1980s. Coherent has been selling Chameleons since…
EM and fluorescence analysis in the same preparation
Posted in Hardware
Several labs have set up their own workflows for imaging the same tissue using both fluorescence and electron microscopy (EM). Zeiss just released a product, called Shuttle-and-Find, to streamline this. It’s pretty simple, just…
Structured illumination
Structured illumination can be used to increase the resolution of wide field fluorescence imaging. The idea is best motivated by thinking about Moire fringes. In panel (a) below, if you stand away from the…
Troubleshooting Fluorescence Microscopy
Posted in Tips
Jim Pawley upgraded the 39 Steps from a simple MacGuffin to a classic checklist for troubleshooting fluorescent microscopy images. Now 10 years old, it remains very relevant. It covers thirty-nine different parameters to…
Measuring the gain of your imaging system
Posted in Hardware
The images obtained from a 2-photon laser scanning microscope, or a CCD-based system, consist of an array of pixel values. For 8-bit…
High power LEDs
Posted in Hardware
About 10 years ago, as high power LEDs became available, many quantitative microscopists started changing over from arc lamps to LEDs for illumination because of their rock solid intensity. Philips Lumileds have been…
Intrinsic imaging tips
Posted in Tips
Intrinsic signal optical imaging is a functional imaging modality where the reflectance of red light indicates active portions of cortex. It is used for many applications, including imaging individual barrels in rodent somatosensory…
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