Live-cell STAR image acquisition was performed with a Nikon Ti-2 microscope equipped with a motorized stage, stage-top incubator to maintain 37 °C and 5% CO2 (Tokai Hit, INUBG2SF-TIZB), ×60 1.49-NA objective, manual TIRF illuminator (Nikon, TI-LA-TIRF), 488 nm (Obis, 488-150 LS), and 647 nm (Obis, 1196627) excitation lasers, fiber coupling optics: fiber mount (Thorlabs, MBT621D), converging and directing the laser objective (Olympus, RMS10X), optical fiber (Thorlabs, P3-405BPM-FC-2), C-NSTORM QUAD 405/488/561/638 nm TIRF dichroic. Images were acquired with an Optosplit III (Cairn Research) image splitter with ET525/50 m and ET705/72 m emission filters (Chroma), and T562lpxtr-UF2 and T640lpxtr-UF2 dichroic mirrors to split the fluorescence emission onto separate regions of the ORCA-Flash 4.0 v3 scientific complementary metal-oxide-semiconductor camera (Hamamatsu). The system was coupled by a data acquisition device (NIDAQ, National Instruments, BNC-2115) and controlled using Nikon Elements software (version 5.02) and Coherent Connection software (version 3.0.0.8). Image acquisition was performed through NIS JOBS. Optosplit III was calibrated using the manufacturer protocol and the NanoGrid (Miraloma Tech, LLC, A00020).
Rms10x
Manufactured by Thorlabs
Sourced in Germany
The RMS10X is a laboratory microscope eyepiece produced by Thorlabs. It has a magnification factor of 10x and a numerical aperture of 0.25. The eyepiece is designed to be compatible with Thorlabs' Rigid Mechanical Standard (RMS) microscope objective mounts.
Automatically generated - may contain errors
2 protocols using rms10x
1
TIRF Microscopy for Live-Cell Imaging
2
Photoluminescence Characterization of Microfluidics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A 375 nm LED (M375L3-Mounted LED, Thorlabs, Germany) was used as
an excitation source for PL measurements. The collimated beam was
directed toward a dichroic beam splitter (Multiphoton LP-Strahlenteiler
HC 375 LP, AHF, Germany) and then focused into the microfluidic channel
using an aspheric lens (A240TM, f = 8.0 mm, NA 0.50, Thorlabs, Germany).
Emission originating from the microfluidic channel was collected by
the same lens, passed through the dichroic beam splitter, and coupled via a 10× objective (RMS10X, NA 0.25, Thorlabs, Germany)
to a fiber spectrometer (QE 65000, Ocean Optics, UK) via a 2 m long multimode fiber with a core diameter of 400 μm
(QP400-2-UV–vis, Ocean Optics, UK). The spectrometer incorporated
a 20 μm entrance slit, a 600 lines/mm grating, and a 2048-pixel
detector. The spectrometer was operated between 350 and 1100 nm, and
data were recorded using an integration time between 50 and 100 ms.
an excitation source for PL measurements. The collimated beam was
directed toward a dichroic beam splitter (Multiphoton LP-Strahlenteiler
HC 375 LP, AHF, Germany) and then focused into the microfluidic channel
using an aspheric lens (A240TM, f = 8.0 mm, NA 0.50, Thorlabs, Germany).
Emission originating from the microfluidic channel was collected by
the same lens, passed through the dichroic beam splitter, and coupled via a 10× objective (RMS10X, NA 0.25, Thorlabs, Germany)
to a fiber spectrometer (QE 65000, Ocean Optics, UK) via a 2 m long multimode fiber with a core diameter of 400 μm
(QP400-2-UV–vis, Ocean Optics, UK). The spectrometer incorporated
a 20 μm entrance slit, a 600 lines/mm grating, and a 2048-pixel
detector. The spectrometer was operated between 350 and 1100 nm, and
data were recorded using an integration time between 50 and 100 ms.
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!