# Technical Mother Machine Core

> **NIH NIH P01** · MASSACHUSETTS EYE AND EAR INFIRMARY · 2022 · $440,396

## Abstract

SUMMARY
The Mother Machine is a microfluidics-based high throughput imaging and analysis system for
measuring the properties and behaviors of single cells in a large population (thousands to millions
of cells). It is capable of identifying rare stochastic variants in a population, such as spontaneous
persister cells, and tracking the properties of those rare cells over time, or in response to various
treatments. It is equally capable of identifying phenotypically-altered genetic variants in a large
pool of mutants, such as a transposon insertion library typically used for high throughput genetic
screens such as Tn-seq. However, rather than relying on indirect inferences based on counts of
sequences of particular transposon insertion junctions following several cycles of PCR
amplification, the Mother Machine allows direct observation of the mutant over time. Moreover, it
allows comparison to about 10 other cells of that genetic lineage in the same trench, providing
extremely rigorous and precise measurements. Additional advantages over conventional Tn-seq
approaches include the ability to identify phenotypes that do not necessary translate to changes
in chromosome abundance (and hence sequence counts), such as the formation of multinucleate
filamentous cells, or other irregularities in cell growth or division. Finally, direct imaging of single
clonal lineages allows direct assessment of the phenotypic consequence for that particular
transposon insertion in that particular orientation, rather than summing the effects of all insertions
across a gene. This allows phenotypic differences that depend on the precise insertion location
of a transposon, and its orientation to be assessed, for individual comparison to other different
insertions within the same gene. The Core Director is a pioneer in this field who has made
advancements in the technology not available anywhere else, including the ability to image more
than 1 million trenches, and the ability to retrieve cells from trenches of interest. This specialized
advanced technology cannot now be recreated in individual laboratories, and is not available
commercially, providing a tremendous, unique resource for the important investigation of antibiotic
resistance and susceptibility of staphylococci and enterococci. Additionally, the Core Director’s
field-leading expertise in stochastic variation within bacterial populations provides an important
intellectual resource for subprojects investigating the properties of antibiotic persister
subpopulations – the effects of antibiotics on them as well as the contribution of variation in efflux
expression to their genesis. These capabilities are highly complementary to those of the research
subprojects, and significantly enhances both their throughput and the rigor and richness of the
data that cannot otherwise be obtained.

## Key facts

- **NIH application ID:** 10327901
- **Project number:** 2P01AI083214-14
- **Recipient organization:** MASSACHUSETTS EYE AND EAR INFIRMARY
- **Principal Investigator:** Johan Paulsson
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $440,396
- **Award type:** 2
- **Project period:** 2009-09-01 → 2026-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10327901

## Citation

> US National Institutes of Health, RePORTER application 10327901, Technical Mother Machine Core (2P01AI083214-14). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10327901. Licensed CC0.

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