Development of Vacuum Ultraviolet Laser Desorption/Ionization Single‑Cell Mass Spectrometry Imaging Instrument

doi:10.19894/j.issn.1000-0518.230360

Chinese Journal of Applied Chemistry ›› 2024, Vol. 41 ›› Issue (1): 100-108.DOI: 10.19894/j.issn.1000-0518.230360

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Development of Vacuum Ultraviolet Laser Desorption/Ionization Single‑Cell Mass Spectrometry Imaging Instrument

Lei XING, Han-Zhang MOU, Jian-Bin PAN(), Bin KANG, Jing-Juan XU(), Hong-Yuan CHEN

State Key Laboratory of Analytical Chemistry for Life Science，School of Chemistry and Chemical Engineering，Nanjing University，Nanjing 210023，China

Received:2023-11-15 Accepted:2023-12-18 Published:2024-01-01 Online:2024-01-30
Contact: Jian-Bin PAN,Jing-Juan XU
About author:xujj@nju.edu.cn
jbpan@nju.edu.cn
Supported by:
the National Natural Science Foundation of China(22034003);the Excellent Research Program of Nanjing University(ZYJH004);the China Postdoctoral Science Foundation(2019TQ0145)

Abstract

Abstract:

Developing the matrix-free mass spectrometry imaging method with both sub-micron resolution and high detection sensitivity is of great significance for visualizing single-cell biological samples. Here， a vacuum ultraviolet laser desorption/ionization time-of-flight mass spectrometry imaging （VUVDI-ToFMSI） method is proposed. By adjusting the frequency wavelengths of three fundamental beams to 777.7， 403.22 and 255 nm， and increasing the argon gas pressure and the temperature in the mercury vapor pool to 1100 Pa and 240 ℃， a high-intensity VUV laser was generated with energy close to 13 μJ/pulse in the laboratory. Meanwhile， the self-designed coaxial optical path focused the VUV laser to the sub-micron level. In addition， by further optimizing the voltage of the deflection， ion focusing， and reflection electric field， the flight path of ions was corrected. Based on this method， nanoscale images （~500 nm/pixel） of exogenous drug methylene blue （m/z 284.4）， endogenous metabolite ions at m/z 152.1 and m/z 81.1 were achieved in a single HeLa cell. The results showed that they were enriched in the cytoplasm， nucleus and nucleolus， respectively， demonstrating the high-resolution and high-detection-sensitivity imaging ability of this method in drawing the distributions of exogenous and endogenous chemicals in single-cell biological samples. The VUVDI-ToFMSI platform will provide more complete and in-depth chemical information for understanding the fine structure and function of more biological samples.

Key words: Single cell, Mass spectrometry imaging, Vacuum ultraviolet, Submicron resolution

CLC Number:

O652

Lei XING, Han-Zhang MOU, Jian-Bin PAN, Bin KANG, Jing-Juan XU, Hong-Yuan CHEN. Development of Vacuum Ultraviolet Laser Desorption/Ionization Single‑Cell Mass Spectrometry Imaging Instrument[J]. Chinese Journal of Applied Chemistry, 2024, 41(1): 100-108.

Figures/Tables 8

Fig.1 Schematic diagram and partial physical images of the VUVDI-ToFMSI device. The instrument mainly includes a system generating the VUV laser （A， B）， a separation chamber （C）， a desorption/ionization chamber （D）， a reflectron tube analyzer （E） and a signal acquisition system

Fig.2 Variation curves of VUV laser intensity with wavelength corresponding to the fundamental lasers at wavelengths of ~403 nm （A） and ~777 nm （B）

Fig.3 Variation curve of VUV laser intensity with （A） argon pressure and （B） the temperature of the mercury vapor

Fig.4 Schematic diagram of screening VUV laser using lens focus method

Fig.5 Changes in VUV laser energy （~13 nJ） during the ~210000 pulses

Fig.6 Changes in mass spectra of PMMA with VR2 voltages （A） and variation curve of the peak intensity with VR2 voltages （B）， located around 38 μs

Table 1 The polarity and optimal values of voltage elative to ground of four adjustable parameters in the reflectron tube of the VUVDI?ToFMSI instrument

Parameters	Polarity	Voltage value/V
Deflection electric field	-	200
Ion focusing lens	+	800
VR1	+	3 500
VR2	+	3 800

Fig.7 （A） Optical image of a single Hela cell. （B） Fluorescence image of DAPI. （C） Fluorescence image of MB. （D） mass image of ions at m/z 152.1. （E） Mass image of fragments of ions at m/z 81.1. （F） Mass image of MB （［C16H18N3S］+ at m/z 284.4）. （G） Merged image of A， B and C. （H） Merged image of D， E and F. All MS images were detected at 500 nm/pixel. Different color brightness represents the ion's abundance and the brighter the color， the stronger the abundance of the ions. Scale bar： 8 μm

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Development of Vacuum Ultraviolet Laser Desorption/Ionization Single‑Cell Mass Spectrometry Imaging Instrument

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