Collection: TIRF Microscopy
The ability to visualize fluorescent molecules with high resolution depends significantly on sample preparation and the optics used, though this can be hindered by out-of-focus light collected within the focal plane.
Total internal reflection fluorescence microscopy (TIRF) utilizes specialized optics to selectively illuminate the sample interface, generating light within a narrow 50-100 nm range. This technique minimizes out-of-focus light, significantly enhancing the detection of fluorescent molecules. TIRF's low-light illumination and exceptional spatial resolution make it invaluable for live-cell imaging applications.
Given its low-light nature, a primary challenge in TIRF microscopy is maximizing the collection of emitted photons to optimize the signal-to-noise ratio. Therefore, employing a highly sensitive camera is essential to capture as many photons as possible, ensuring accurate and clear imaging results in live-cell studies.
