Imaging was conducted on an Olympus FV1200MPE BASIC (BX-61WI) microscope equipped with a 25×, 1.05 NA water-immersion objective (XLPL25XWMP, Olympus) and an ultrafast pulsed laser (Mai Tai DeepSee HP, Spectra-Physics) tuned to 910 nm. Epifluorescence emission was separated into “green” and “red” channels with a 570nm dichroic mirror and a 525/50 bandpass filter (FF03-525/50-32, Semrock, green channel) and 575-630nm bandpass filter (BA575-630, Olympus, red channel), respectively. The microscope system was controlled by FluoView software (FV10-ASW v.4.1). Images of 256 × 128 pixels representing 256 × 128 μm on the retina were acquired at 15 Hz (zoom setting of 2).
Fv1200mpe basic bx 61wi microscope
Manufactured by Olympus
The FV1200MPE BASIC (BX-61WI) is a high-performance upright microscope system designed for advanced imaging applications. It features a BX-61WI inverted stand with a motorized XY stage and Z-drive. The microscope is capable of performing multi-photon excitation imaging.
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3 protocols using fv1200mpe basic bx 61wi microscope
1
Two-Photon Imaging of Retinal Specimens
2
Fluorescent Cell Targeting for Patch Recordings
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To target fluorescent cells for patch recording, two photon imaging was used (Olympus FV1200MPE BASIC (BX-61WI) microscope, 25×, 1.05 NA water-immersion objective (XLPL25XWMP, Olympus), an ultrafast pulsed laser (Mai Tai DeepSee HP, Spectra-Physics) tuned to 910 nm, and the imaging software Fluoview 4.1 (Olympus)). To acquire an image stack, RGCs were filled during electrophysiological recordings with Alexa hydrazide 488 or 594 (100 µM, Invitrogen), and were imaged following the recording, either using the two-photon or the single-photon (confocal) configuration of the two-photon microscope. Tissue in which fluorescent proteins were expressed was often fixed and immunostained (see above), and subsequently imaged on a confocal microscope (Olympus FV3000, UPlan Super Apochromat objectives, 30xS, 1.05 NA, or 60x2S, 1.3 NA) in the Leduc Imaging Facility, Brown University. Confocal and two-photon stacks were processed in Fiji (https://imagej.net/software/fiji ), and collapsed using either maximum intensity or maximum standard deviation projections.
3
Two-Photon Imaging of Retinal Specimens
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Imaging was conducted on an Olympus FV1200MPE BASIC (BX-61WI) microscope equipped with a 25×, 1.05 NA water-immersion objective (XLPL25XWMP, Olympus) and an ultrafast pulsed laser (Mai Tai DeepSee HP, Spectra-Physics) tuned to 910 nm. Epifluorescence emission was separated into “green” and “red” channels with a 570nm dichroic mirror and a 525/50 bandpass filter (FF03-525/50-32, Semrock, green channel) and 575-630nm bandpass filter (BA575-630, Olympus, red channel), respectively. The microscope system was controlled by FluoView software (FV10-ASW v.4.1). Images of 256 × 128 pixels representing 256 × 128 μm on the retina were acquired at 15 Hz (zoom setting of 2).
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