MALDI MS Imaging on the 4800 MALDI TOFTOF
- Slides: 26
MALDI MS Imaging on the 4800 MALDI TOF/TOF™ Analyzer Prepared by Andrew James, Ph. D
MALDI Imaging Mass Spectrometry (IMS) • The analysis of a sample surface for its molecular content – Sample consists of a thinly sliced section of tissue/organ/whole animal • A 2 dimensional array of MALDI spectra are obtained over the surface of the sample – each spectra has location component • An ion intensity map can then be produced for any mass that is detected over the scanned area.
Sample Preparation • Coronal Rat Brain Sections – 12 um thickness • After sample mounting, tissue sections were washed in 70% Et. OH solution for 30 sec – Then dried under vacuum • Tissue sections were spray coated with CHCA (10 mg/m. L 60% Acn, 40%water, 0. 1%TFA) – Approx. 15 coats
Image Acquisition • 4800 data • MSI imaging acquisition tool and 4000 Series Explorer used together Setup • Laser: 50 shots/pixel • Pixel size 200 um x 200 um • Linear TOF acquisition method: 3 -25 k. Da (note: No active processing method)
Slice Tissue Sections Mount Sample Data Processing Spray Coat with Matrix 4800 MALDI TOF/TOF Load Sample Acquisition Software
Image Analysis • Image analysis was carried out on Bio. Map software – Produce Images – Data manipulation (baseline correction on underlying spectra) – Compare results images
Active Image file Optical Image
Active Image Window MS Image result windows Optical Image
• Active image window – Contains all the spectral information – Use to generate specific MS images • MS image results windows – Intensity map image for a specific mass (protein or bio-molecule) • Optical images – Scanned image of sample prior to matrix application
Relative Intensity
Image Overlay
Regions Of Interest (ROI) • Compare Spectra from 2 different areas of tissue sample • Compare intensities of a given compound in 2 different areas of tissue
Comparing Spectra in Different ROIs • Draw ROI in the Active Image data window
Compare average spectra in different regions Define ROIs
Compare Spectra from multiple ROIs
• Useful for visualizing differences • Can drag cursor over the over-layed spectra to generate images in the active image data window
Comparing Intensity of an Analyte of Interest • Draw ROIs within an MS image result window
Compare intensity of detection protein in specific regions of sample
Compare average intensity of a detected protein in specific regions of the sample Average intensity of detected protein within the specified ROI
• Get statistics for average intensity of a specific mass in different regions of the tissue section – Relative quantitation
Conclusion • Still some work to do with sample preparation – Should be seeing more complex spectra – Compare spotting vs. spraying of matrix – Surface charge affects? • Data acquisition may be improved by using an rastering pattern instead of a stationary shot pattern over the sample • Biomap is a very good processing tool
- Esi and maldi
- "green imaging" -g -"green imaging technologies"
- Early beginnings of tourism
- Sistema vds
- 4800 oak grove drive pasadena
- 61000x12
- 4800 oak grove drive
- Mazak fh 4800
- Diễn thế sinh thái là
- Vẽ hình chiếu vuông góc của vật thể sau
- Giọng cùng tên là
- Làm thế nào để 102-1=99
- Tỉ lệ cơ thể trẻ em
- Lời thề hippocrates
- Hổ đẻ mỗi lứa mấy con
- đại từ thay thế
- Quá trình desamine hóa có thể tạo ra
- Công của trọng lực
- Thế nào là mạng điện lắp đặt kiểu nổi
- Hát kết hợp bộ gõ cơ thể
- Các loại đột biến cấu trúc nhiễm sắc thể
- Vẽ hình chiếu đứng bằng cạnh của vật thể
- Biện pháp chống mỏi cơ
- độ dài liên kết
- Chó sói
- Môn thể thao bắt đầu bằng từ chạy
- Khi nào hổ con có thể sống độc lập