Fiduciary-Free Frame Alignment for Robust Time-Lapse Drift Correction Estimation in Multi-Sample Cell Microscopy

TL;DR

This paper introduces a new method for aligning microscope images to correct drift and jitter in multi-sample cell observations.

Contribution

A robust, sub-pixel precise frame alignment method using RAFT for drift correction in multi-sample microscopy.

Findings

The RAFT model achieved near-perfect precision in aligning time-lapse frames across ten unrelated datasets.

The method is robust for elastically undistorted and translation-displaced microscopic observations.

The approach works well for registration but not for tracking individual image components like cells.

Abstract

When analyzing microscopic time-lapse observations, frame alignment is an essential task to visually understand the morphological and translation dynamics of cells and tissue. While in traditional single-sample microscopy, the region of interest (RoI) is fixed, multi-sample microscopy often uses a single microscope that scans multiple samples over a long period of time by laterally relocating the sample stage. Hence, the relocation of the optics induces a statistical RoI offset and can introduce jitter as well as drift, which results in a misaligned RoI for each sample’s time-lapse observation (stage drift). We introduce a robust approach to automatically align all frames within a time-lapse observation and compensate for frame drift. In this study, we present a sub-pixel precise alignment approach based on recurrent all-pairs field transforms (RAFT); a deep network architecture for optical flow. We show that the RAFT model pre-trained on the Sintel dataset performed with near perfect precision for registration tasks on a set of ten contextually unrelated time-lapse observations containing 250 frames each. Our approach is robust for elastically undistorted and translation displaced (x,y) microscopic time-lapse observations and was tested on multiple samples with varying cell density, obtained using different devices. The approach only performed well for registration and not for tracking of the individual image components like cells and contaminants. We provide an open-source command-line application that corrects for stage drift and jitter.

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