Single Cell Analysis with the MVX-7100 L Workstation July 17 th 2019 - - PowerPoint PPT Presentation

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Single Cell Analysis with the MVX-7100 L Workstation July 17 th 2019 - - PowerPoint PPT Presentation

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Single Cell Analysis with the MVX-7100 L Workstation July 17 th 2019 Peter Winship, Ph.D. Webinar Agenda An introduction to cell analysis with ICP-MS Webinar Agenda An introduction to cell analysis with ICP-MS The single cell

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SLIDE 1

Single Cell Analysis with the MVX-7100 µL Workstation

July 17th 2019 Peter Winship, Ph.D.

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SLIDE 2

Webinar Agenda

  • An introduction to cell analysis with ICP-MS
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SLIDE 3

Webinar Agenda

  • An introduction to cell analysis with ICP-MS
  • The single cell analysis
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SLIDE 4

Webinar Agenda

  • An introduction to cell analysis with ICP-MS
  • The single cell analysis
  • From sample to data: the single cell workflow

– Automation – Glassware interface – ICP-MS

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SLIDE 5

Webinar Agenda

  • An introduction to cell analysis with ICP-MS
  • The single cell analysis
  • From sample to data: the single cell workflow

– Automation – Glassware interface – ICP-MS

  • Wider analytical considerations
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SLIDE 6

Cell Analysis and ICP-MS

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SLIDE 7

Cell Analysis and ICP-MS

  • The latest generation of ICP-MS affords the

sensitivity and speed to detect ‘single events’

  • Huge area of research and analysis in the

elemental composition of eukaryotic and prokaryotic cells

– Elemental content of individual cells – Cellular uptake of species (nutrient/contaminant) – Uptake of metal based drug compounds, nanoparticles

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SLIDE 8

The Single Cell Analysis

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SLIDE 9

The Single Cell Analysis

  • Dissolved analyte: continuous sample introduction
  • Suspension of cells: individual cells produce an ion plume

Time Intensity Dwell time Time Intensity Dwell time

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SLIDE 10

The Single Cell Analysis

  • Individually introduced cells

produce an ion plume in plasma

  • Number of events equivalent to cell

number concentrations

  • Signal intensity of events equivalent

to element mass in individual cells

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SLIDE 11

From Sample to Data: The Single Cell Workflow

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SLIDE 12

The Analytical Workflow

Sample Automation Interface ICP-MS Data

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SLIDE 13

The Analytical Workflow

Sample Automation Interface ICP-MS Data

  • Transport Efficiency: Maximum
  • Maintain Cell Integrity
  • Numerous variables
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SLIDE 14

Sample

  • Consider the cell processes prior to analysis
  • Samples delivered in certain media
  • Samples in well plates/tubes/vials
  • Are there conditions to maintain?

– E.g. temperature?

  • Any other considerations?

– Cell gravitational settling?

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SLIDE 15

Automation

  • MVX-7100 µL Workstation

– Syringe driven automation – Sample volume: 5 µL to 1.5 mL – Control of introduction flow rate (nL min-1 to mL min-1) – Sample homogenization – Temperature control (4°C to 40°C) – Sampling from vials/tubes – Sampling from 96 or 384 well plates

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SLIDE 16

Glassware Interface: Glass Expansion

  • High efficiency nebulizer
  • Introduction flow rates < 50 µL min-1
  • Wide bore capillary
  • On axis, high efficiency spray chamber
  • Additional spray chamber gas flow

focusses nebulized sample

  • ‘Near total consumption’
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SLIDE 17

Glassware Interface: Glass Expansion

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SLIDE 18

Confirmation of Cell Integrity

  • K562 cell line

– Human Myeloid Leukemia cells – 10 to 20 µm size – Widely used in research – Easy to culture and isolate

Images courtesy of Agilent Technologies

20 µm

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SLIDE 19

Confirmation of Cell Integrity

Post nebulisation Tris-HCl 50 mM MVX-7100 µL Workstation Sample Loop and 6 Port Switching Valve Intact Intact Lysis Nebuliser Gas Flow = 0.1 L min-1 Nebuliser Gas Flow = 0.7 L min-1

Images courtesy of Agilent Technologies

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SLIDE 20

ICP-MS Instrumentation

  • Analytical sensitivity

– The more sensitivity available the better – Broad range of associated analyte concentrations

  • Analyte suite?

– Multiple analytes: the analyte of interest and a co-factor? – Full suite from ICP-TOF-MS

  • Data processing

– What is the data output? – Concentration/population number/cell size? – Analyte per unit x?

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SLIDE 21

Data Output Example

31P+>31P16O+ 80Se+>80Se16O+ 31P+>31P16O+ 80Se+>80Se16O+

Selenised Yeast Regular Yeast

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SLIDE 22

Wider Analytical Considerations

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Wider Considerations and Further Work

  • Standardisation

– How do we standardize? – Cell size/cell shape/cell integrity

  • Quality control
  • Interlaboratory comparison

– Standardisation and QC testing

  • High TDS testing
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SLIDE 24

Acknowledgements

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SLIDE 25

Acknowledgements

  • Agilent Technologies
  • Glyn Russell, Glass Expansion
  • Jorg Bettmer, University of Oviedo, Spain
  • Dr Tanaka and Prof. Ogra, Chiba University, Japan
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SLIDE 26

Thank You For Your Attention

Please forward questions to pete.winship@teledyne.com

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SLIDE 27

www.teledynecetac.com

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SLIDE 28

Peter Winship, Ph.D MVX-7100 µL Workstation Product Manager pete.winship@teledyne.com