Use of IR spectroscopy to evaluate penetration of reagents into biological specimen
Inventors
Shah, Amit D. • Flores, Cristina R.
Assignees
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Publication Number
US-11460400-B2
Publication Date
2022-10-04
Expiration Date
Abstract
Apparatuses for measuring sample materials (such as tissue samples) and associated methodologies and instrumentation involve a spectroscopy system or device configured with a sample measuring device (e.g., an attenuated total reflection infrared (ATR-IR) sample measuring device) including a crystal defining a sample receiving surface/interface operatively connected to an IR source and an IR detector. The sample receiving surface/interface facilitates assessing presence (and optionally, amount) of a fixative (e.g., formaldehyde) in sample materials, tissue samples for example. Measurements are taken at one or multiple locations within a sample placement structure/area, which can be provided in the form of a probe or a raised piercing structure containing one or more ATR-IR (or other) sample measuring devices. The probe can include a piercing structure or mechanism at a distal (open) end of the probe configured to create a passage to a selected location (depth) within the sample material in a minimally invasive manner so that the sample is not severely damaged during testing.
Core Innovation
The disclosure relates to an attenuated total reflection infrared (ATR-IR) sample material measuring method and system for evaluating penetration of a fixative substance into a sample material. An spectroscopy system or device is configured with a probe including an ATR-IR sample measuring device having a crystal defining a sample receiving surface/interface operatively connected to an IR source and an IR detector.
The IR source generates an internally reflected IR beam propagating through the crystal while the material at a selected depth location is against the sample receiving surface/interface. The IR detector records an attenuated IR beam output of the crystal, and the attenuated IR beam is used to generate/provide an IR spectrum or IR spectra. The IR spectrum(s) are evaluated to determine whether a signal is present indicating a presence of a fixative substance in the sample material at the selected depth location.
The disclosure specifically addresses formaldehyde-related IR absorption, including IR absorption associated with C=O stretch in the range of about 1000 cm−1 to 1100 cm−1. Spatial selection and repeat measurements are supported by advancing or placing the probe so that material at a selected depth location is brought directly into contact with the sample receiving surface/interface at one or multiple locations, with a repositionable probe structure and an ATR crystal arrangement for measurements at multiple locations using single or multiple ATR-IR crystals.
Claims Coverage
The independent claims are clm-00001, clm-00008, and clm-00014, covering three inventive features.
Selected-depth ATR-IR spectrum evaluation for fixative signal
An spectroscopy system or device with a probe containing an ATR-IR sample measuring device including a crystal defining a sample receiving surface/interface operatively connected to an IR source and an IR detector; advancing the probe into a sample bringing material at a selected depth location directly into contact with the sample receiving surface/interface; generating an internally reflected IR beam propagating through the crystal; recording an attenuated IR beam output; generating/providing an IR spectrum or IR spectra; and evaluating the IR spectrum/spectra to determine whether a signal is present indicating a presence of a fixative substance in the sample material at the selected depth location.
Repositionable ATR-IR sample measuring device in a probe apparatus
A spectroscopy system or device comprising a probe comprising an attenuated total reflection infrared (ATR-IR) sample measuring device comprising a crystal defining a sample receiving surface/interface, wherein the sample measuring device is configured to be repositionable between a forward/extended position adjacent a distal end of the probe and a rearward/retracted position recessed within the probe enclosure away from the distal end.
ATR-IR sample contact and spectrum evaluation for fixative presence
An spectroscopy system or device with an ATR-IR sample measuring device including a crystal defining a sample receiving surface/interface operatively connected to an IR source and an IR detector; placing a sample material on the ATR-IR sample measuring device bringing the sample material directly into contact with the sample receiving surface/interface; generating an internally reflected IR beam propagating through the crystal; recording an attenuated IR beam output; generating/providing an IR spectrum or IR spectra; and evaluating the IR spectrum/spectra to determine whether a signal is present indicating a presence of a fixative substance in the sample material.
Across the independent claims, the core coverage is ATR-IR spectroscopy using an ATR crystal with an IR source and IR detector to generate an IR spectrum from an attenuated IR beam and evaluate whether a signal indicating presence of a fixative substance is present, with selected depth location evaluation, a repositionable ATR-IR measuring device in a probe, and sample contact with the ATR surface.
Stated Advantages
Enables determination of whether a signal is present indicating a presence of a fixative substance in the sample material at a selected depth location.
Supports qualitative go/no-go and optional quantitative estimation based on formaldehyde-related IR absorption.
Allows visualization of spectra patterns and results from measurements taken at different times and/or locations.
Documented Applications
Evaluating penetration of fixatives into biological specimens, including evaluating penetration of formalin/formaldehyde into tissue samples at selected depth locations and supporting go/no-go and optional quantitative estimation.
Measuring sample material at one or multiple spatial locations and obtaining measurements at different times, including monitoring IR spectral features that grow during infiltration over time.
Processing and visualization of ATR-IR spectra (e.g., FTIR spectra) with indicators and a user interface for displaying analysis status based on evaluated signals.
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