# Spectroscopic Approach to Correction and Visualisation of Bright-Field Light Transmission Microscopy Biological Data

## TL;DR

This paper introduces a spectroscopic correction method for bright-field microscopy data that improves image accuracy by calibrating optical path distortions, enabling better visualization of biological samples like live cells.

## Contribution

A novel spectroscopic approach for correcting and visualizing bright-field microscopy data by calibrating optical distortions and sensor sensitivities.

## Key findings

- Corrected 12-bit images accurately reflect photon flux.
- Enhanced visualization with minimal information loss.
- Method improves data fidelity in biological microscopy.

## Abstract

The most realistic information about the transparent sample such as a live cell can be obtained only using bright-field light microscopy. At high-intensity pulsing LED illumination, we captured a primary 12-bit-per-channel (bpc) response from an observed sample using a bright-field microscope equipped with a high-resolution (4872x3248) image sensor. In order to suppress data distortions originating from the light interactions with elements in the optical path, poor sensor reproduction (geometrical defects of the camera sensor and some peculiarities of sensor sensitivity), we propose a spectroscopic approach for the correction of this uncompressed 12-bpc data by simultaneous calibration of all parts of the experimental arrangement. Moreover, the final intensities of the corrected images are proportional to the photon fluxes detected by a camera sensor. It can be visualized in 8-bpc intensity depth after the Least Information Loss compression.