Cells were seeded at a density of 1 × 103 per well in 96 well 1.5 glass bottom plates (Cellvis) coated with 10 μg/ml Fibronectin (Huber lab) and incubated for 24 h at 37°C with 5% CO2. Before imaging, cells were washed with PBS and incubated in starving medium for 4 h in the dark. Imaging was performed with an epifluorescence Eclipse Ti inverted fluorescence microscope (Nikon) using a CFI Apochromat TIRF 100× oil (NA 1.49). Images were acquired with an Andor Zyla 4.2 plus camera at a 16‐bit depth. TIRF images were acquired with a 561 nm laser using a ET575/25 filter in front of the ZT488/561rpc (Chroma) to prevent nonspecific activation of the CRY2. MetaMorph software (Universal Imaging) was used for acquisition. TIRF images of the optoFGFR‐mScarlet were acquired at a 20‐s interval. Optogenetic stimulation was done using a 470 nm LED (SPECTRA X, Lumencor) (Appendix Fig S1B ). All experiments were carried on at 37°C with 5% CO2.
Cfi apochromat tirf 100 oil
Manufactured by Nikon
Sourced in Japan
The CFI Apochromat TIRF 100× oil is a high-magnification objective lens designed for Total Internal Reflection Fluorescence (TIRF) microscopy. It features an apochromatic correction to minimize chromatic aberration and provide superior optical performance.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse ti, by Nikon (1 mentions)
Spectra x, by Lumencor (1 mentions)
Metamorph software, by Universal Imaging (1 mentions)
Nis element, by Nikon (1 mentions)
Goat serum, by Thermo Fisher Scientific (1 mentions)
Prolong glass antifade, by Thermo Fisher Scientific (1 mentions)
Cfi plan apochromat lambda 20, by Nikon (1 mentions)
A1r hd confocal microscope, by Nikon (1 mentions)
2 protocols using cfi apochromat tirf 100 oil
1
Optogeentic FGFR Signaling in Cells
2
Immunofluorescence Labeling of Primary Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary neurons on cover glasses or 20 μm cryostat-sectioned coronal brain slices were incubated for 1 h at room temperature with blocking buffer (PBS containing 5% [v/v] goat serum [catalog no.: 16210-064; Gibco, New Zealand], 1% [w/v] bovine serum albumin, and 0.3% [v/v] Triton X-100), followed by 2 h of incubation with primary antibodies (Table 1 ). After washing in PBS, the samples were incubated with AlexaFluor-conjugated secondary antibodies, washed again, mounted for microscopy with ProLong Glass Antifade (catalog no.: P36980; Invitrogen), and examined under a Nikon A1RHD confocal microscope with a CFI Apochromat TIRF 100× Oil, numerical aperture 1.49 or CFI Plan Apochromat Lambda 20×, numerical aperture 0.75 (Nikon Instruments, Tokyo, Japan). Images were acquired with NIS-elements (Laboratory Imaging, Nikon) and then processed in ImageJ (NIH, Bethesda, MD).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!