Rapid detection of replicating cells

摘要

The invention enables efficient, rapid, and sensitive enumeration of living cells by detecting microscopic colonies derived from in situ cell division using large area imaging. Microbial enumeration tests based on the invention address an important problem in clinical and industrial microbiology—the long time needed for detection in traditional tests—while retaining key advantages of the traditional methods based on microbial culture. Embodiments of the invention include non-destructive aseptic methods for detecting cellular microcolonies without labeling reagents. These methods allow for the generation of pure cultures which can be used for microbial identification and determination of antimicrobial resistance.

主权项

1. An instrument for detecting microcolonies of target cells in a sample, said instrument comprising:
(a) a photoelectric array detector having associated optics to detect a detection area having at least one dimension that is ≧1 cm without magnifying said detection area by more than 5-fold and an optical resolution of less than 50 microns; (b) an illumination source that illuminates said detection area; and (c) a computer programmed to receive data collected by said photoelectric array detector, wherein said data is a digital representation of said detection area, and programmed for image analysis comprising analyzing said data to detect one or more microcolonies having a measurement of less than 50 microns in at least two orthogonal dimensions, and programmed to quantify the number of microcolonies detected in said detection area,wherein said instrument detects a property of said one or more microcolonies that does not depend on the addition of a signaling moiety or category binding molecule, and wherein said cells in said one or more microcolonies remain competent to replicate following detection.