High speed modulation sample imaging apparatus and method
AssigneeFluidigm Canada Inc.
InventorsDAAF SANDKUIJL, Alexander V. Loboda, ADAM CAREW, MINALINI LAKSHMAN
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Abstract
This disclosure relates to systems and methods for high speed modulation sample imaging. Disclosed herein are systems and methods for performing imaging mass cytometry, including analysis of labelling atoms by elemental (e.g., atomic) mass spectrometry. Aspects include a sampling system having, and method of using, a femtosecond (fs) laser and/or laser scanning. Alternatively or in addition, aspects include systems and methods for co-registering other imaging modalities with imaging mass cytometry.
Claims69
An apparatus for analysing a biological sample, comprising:
- (i) a sampling and ionisation system to remove material from the sample and to ionise said material to form elemental ions, comprising a laser source, a laser scanning system and a sample stage.
The apparatus according to any preceding claim wherein the laser scanning system response time is quicker than 1 ms, quicker than 500 μs, quicker than 250 μs, quicker than 100 μs, quicker than 50 μs, quicker than 10 μs, quicker than 5 μs, quicker than 1 μs, quicker than 500 ns, quicker than 250 ns, quicker than 100 ns, quicker than 50 ns, quicker than 10 ns, or around 1 ns.
The apparatus of any preceding claim wherein the laser source is a picosecond laser or a femtosecond laser, in particular a femtosecond laser, optionally comprising a pulse picker, such as wherein the pulse picker is controlled by a control module that also controls the movement of the sample stage and/or the positioner(s) of the laser scanning system.
The apparatus of any preceding claim wherein:
- (i) the ablation rate is 200 Hz or greater, such as 500 Hz or greater, 750 Hz or greater, 1 kHz or greater, 1.5 kHz or greater, 2 kHz or greater, 2.5 kHz or greater, 3 kHz or greater, 3.5 kHz or greater, 4 kHz or greater, 4.5 kHz or greater, 5 kHz or greater, or 10 kHz or greater, around 100 kHz, 100 kHz or greater, 1 MHz or greater, 10 MHz or greater, or 100 MHz or greater;
- and/or
- (ii) the laser repetition rate is at least 1 kHz, such as at least 10 kHz, at least 100 kHz, at least 1 MHz, at least 10 MHz, around 50 MHz, or at least 100 MHz, optionally wherein the sampling system further comprises a pulse picker, such as wherein the pulse picker is controlled by a control module that also controls the movement of the sample stage and/or the positioner(s) of the laser scanning system.
The apparatus of any preceding claim wherein the laser source is adapted to produce a spot size of diameter less than 10 μm, less than 5 μm, less than 2 μm, around 1 μm, or less than 1 μm.
The apparatus according to any preceding claim further comprising a camera.
The apparatus according to any preceding claim in which the ionisation system is an ICP.
The apparatus according to any preceding claim in which the detector is a TOF mass spectrometer.
A method of analysing a sample comprising:
- (i) performing laser ablation of the sample on a sample stage, wherein laser radiation is directed onto the sample using a laser scanning system, and wherein the ablation is performed at multiple locations to form a plurality of plumes;
- and
- (ii) subjecting the plumes to ionisation and mass spectrometry, whereby detection of atoms in the plumes permits construction of an image of the sample, optionally wherein the multiple locations are multiple known locations.
A method of performing mass cytometry on a sample comprising a plurality of cells, the method comprising:
- (i) labelling a plurality of different target molecules in the sample with one or more different labelling atoms, to provide a labelled sample;
- (ii) performing laser ablation of the sample on a sample stage, wherein laser radiation is directed onto the sample using a laser scanning system, and wherein the ablation is performed at multiple locations to form a plurality of plumes;
- and
- (iii) subjecting the plumes to ionisation and mass spectrometry, whereby detection of atoms in the plumes permits construction of an image of the sample, optionally wherein the multiple locations are multiple known locations.
A method of analysing a sample comprising:
- desorbing a slug of sample material using laser radiation, wherein laser radiation is directed onto the sample on a sample stage using a laser scanning system;
- and
- (ii) ionising the slug of sample material and detecting atoms in the slug by mass spectrometry.
A method of performing mass cytometry on a sample comprising a plurality of cells, the method comprising:
- (i) labelling a plurality of different target molecules in the sample with one or more different labelling atoms, to provide a labelled sample;
- (ii) desorbing a slug of sample material using laser radiation, wherein laser radiation is directed onto the sample on a sample stage using a laser scanning system;
- and
- (iii) ionising the slug of sample material and detecting atoms in the slug by mass spectrometry.
A laser scanning system for use in any one of methods 22-57.
A method of coregistering images, comprising:
- a) Obtaining a first image from a first tissue section of a tissue sample by an imaging modality other than imaging mass cytometry;
- b) Obtaining a second image of a second tissue section of the tissue sample by imaging mass cytometry;
- c) Coregistering the first and second images.
A method of imaging mass cytometry comprising;
- Identifying a feature in a sample by optical microscopy;
- Scanning radiation across that feature to produce a plume of material;
- Delivering the plume of material to a mass analyser.
Dependencies are inferred from the claim text.