Multi-camera Simultaneous Total Internal Reflection and Interference Reflection Microscopy DOI Creative Commons

Jeffrey O. Spector,

Jiayi Chen, Antonina Roll‐Mecak

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Авг. 28, 2024

Abstract Interference Reflection Microscopy (IRM) is an optical technique that relies on the interference between reflected light from incident beam as it passes through materials of different refractive indices. This has been successfully used to image microtubules, biologically important biofilaments with a diameter 25 nm. However, often desirable both microtubule and interacting proteins simultaneously. Here we present simple modification standard multi-color total internal reflection fluorescence (TIRF) microscope enables simultaneous high-speed IRM single molecule TIRF imaging. Our design utilizes camera for each channel (IRM TIRF) allowing independent optimization parameters two modalities. We illustrate its application by imaging unlabeled microtubules GFP-labeled end-binding protein EB1 which forms comets tips polymerizing microtubules. easily implemented, minimal cost, making accessible any laboratory existing microscope.

Язык: Английский

Multi-camera Simultaneous Total Internal Reflection and Interference Reflection Microscopy DOI Creative Commons

Jeffrey O. Spector,

Jiayi Chen, Antonina Roll‐Mecak

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Авг. 28, 2024

Abstract Interference Reflection Microscopy (IRM) is an optical technique that relies on the interference between reflected light from incident beam as it passes through materials of different refractive indices. This has been successfully used to image microtubules, biologically important biofilaments with a diameter 25 nm. However, often desirable both microtubule and interacting proteins simultaneously. Here we present simple modification standard multi-color total internal reflection fluorescence (TIRF) microscope enables simultaneous high-speed IRM single molecule TIRF imaging. Our design utilizes camera for each channel (IRM TIRF) allowing independent optimization parameters two modalities. We illustrate its application by imaging unlabeled microtubules GFP-labeled end-binding protein EB1 which forms comets tips polymerizing microtubules. easily implemented, minimal cost, making accessible any laboratory existing microscope.

Язык: Английский

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