To identify cells within each well, we rst imaged the device via bright-eld imaging with 2x2 binning using a 4x objective. To identify embedded spectral codes within each bead, we then illuminated the device from above using 292 nm excitation via a Xenon arc lamp (Lambda LS, Sutter Instruments, Novato, CA) equipped with an automated lter wheel (Lambda 10-2, Sutter Instruments, Novato, CA) containing a 292/27 bandpass excitation lter (Semrock, Rochester, NY) paired with UG11 absorptive glass (Newport, Irvine, CA). Emitted light was passed through an additional UV blocking lter mounted within a custom 3D printed holder mounted over the objective and then collected within Ln-channels using nine emission lters (435/40, 474/10, 536/40, 546/6, 572/15, 620/14, 630/92, 650/13, and 780/20 nm). For each image, we then identi ed all beads and determined the Ln ratios most likely to have produced the observed spectra associated with each pixel via linear unmixing relative to a series of Ln reference spectra as described previously 7 . Finally, we created a matrix associating each microwell (indexed by row and column) with the spectral code of the bead within it.
Bandpass excitation lter
Manufactured by IDEX Corporation
The 292/27 bandpass excitation filter is a lab equipment product designed to selectively pass a specific range of frequencies while attenuating frequencies outside that range. It functions as a filter, allowing the desired frequency band to pass through while blocking unwanted frequencies.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using bandpass excitation lter
1
Spectral Barcoding for Single-Cell Identification
2
Spectral Barcoding for Single-Cell Identification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To identify cells within each well, we rst imaged the device via bright-eld imaging with 2x2 binning using a 4x objective. To identify embedded spectral codes within each bead, we then illuminated the device from above using 292 nm excitation via a Xenon arc lamp (Lambda LS, Sutter Instruments, Novato, CA) equipped with an automated lter wheel (Lambda 10-2, Sutter Instruments, Novato, CA) containing a 292/27 bandpass excitation lter (Semrock, Rochester, NY) paired with UG11 absorptive glass (Newport, Irvine, CA). Emitted light was passed through an additional UV blocking lter mounted within a custom 3D printed holder mounted over the objective and then collected within Ln-channels using nine emission lters (435/40, 474/10, 536/40, 546/6, 572/15, 620/14, 630/92, 650/13, and 780/20 nm). For each image, we then identi ed all beads and determined the Ln ratios most likely to have produced the observed spectra associated with each pixel via linear unmixing relative to a series of Ln reference spectra as described previously 7 . Finally, we created a matrix associating each microwell (indexed by row and column) with the spectral code of the bead within it.
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!