MasSpec Pen could potentially be used as a clinical and intraoperative technology for ex vivo and in vivo cancer diagnosis.

1. Nondestructive Tissue Analysis
For Ex Vivo And In Vivo Cancer Diagnosis
Using A Handheld Mass Spectrometry
System
Name: Nalesh Jadhav
Roll no: 201707018
1
Zhang et al., Sci. Transl. Med. 9, eaan3968 (2017)
6 September 2017
PMID: 28878011
DOI: 10.1126/scitranslmed.aan3968

2. What is Cancer?
2
Normal Cells
Cells With
Damaged DNA
Cell Division
Eventually Stops
Cell Division
Never Stops

3. How Cancer Begins?
CancerInfections
Environmental
Factors
Inherited
Genetic
Defects
Poor Lifestyle
Choices
Accumulation
Of Defects
Mutations
3https://www.cancer.gov/about-cancer/diagnosis-staging/diagnosis

4. How is Cancer Usually Diagnosed?
■ Physical Examination: Doctor may
feel areas of your body for lumps that
are formed due to formation of tumor
that may indicate a tumor.
4
■ Laboratory Tests: Urine and blood
tests may help doctor identify
abnormalities that can be caused by
cancer

5. ■ Imaging Tests: Examine bones and internal
organs in a noninvasive way.
– (CT) scan, bone scan,
– magnetic resonance imaging (MRI),
– X-ray
5
■ During a BIOPSY, Doctor collects a sample of
cells for testing in the laboratory. There are
several ways of collecting a sample
How is Cancer Usually Diagnosed?

6. Frozen section preparation is
time- and labor-intensive and
requires skilled technicians and
pathologists to produce and
interpret the results
Freezing artifacts can
negatively interfere with tissue
structure and cell morphology,
thus complicating pathological
interpretation
Intraoperative frozen section
analysis prolongs operative
time, subjecting the patient to
increased risks related to
extended anesthesia
Additional surgical procedures
for re-excision of the involved
margin, which increases health
care costs and places the
patient at risk for additional
surgical complications,
discomfort, and anxiety
6
What are the Drawbacks of these
Methods?

7. How is Molecular Diagnostics Done?
7
Microscopic
Analysis
Patient’s
Tissue Sample
Pathology
Analysis of cell morphology

8. How is Molecular Diagnostics Done?
8
Proteomic
Analysis
Patient’s
Tissue Sample
Proteomic
image
Mass Spectra analysis

9. ■ It is a instrumental technique in which sample is
converted to rapidly moving charged ions by electron
bombardment and they are separated according to their
masses
■ Molecular ions formed in the source acquire some kinetic
energy and leave the source
■ Then the passing ions are analyzed as a function of their
mass to charge ratios
9

10. What Kinds of Sample can be Chosen for
Mass Spec?
10
ExVivo Diagnosis
• Frozen Samples
• Fresh Samples
InVivo Diagnosis
• Live Organism

11. How was Mass Spec used to Detect
Cancer Previously?
11
Rapid Evaporative Ionization Mass Spectrometry – Intelligent Knife

12. ■ At the best-equipped hospitals current surgical practices
may take at least 10–30 minutes for Detection related to
tumor tissue or organ removal
■ Tissue is cut through a electron spray which ionizes the
sample and damages the tissue
■ Surgeons are not able to determine during surgery
whether every cancer cell and tissue has been removed
■ If cancer cells remain after surgery, the tumor tissue may
require further surgery
12
Drawbacks of IKNIFE

13. Aim Of The Study
■ Nondestructive Tissue Analysis by using Mass
Spectrometry System
■ Reduce the Time for cancer diagnosis
■ Reduce cost of sample prep for mass spec
analysis
■ To design a technique that will detect cancer
intraoperatively
13

14. Flow Of The Work
14
Designed
Mass Spec
Pen
Tested Ex
Vivo
Samples
Tested In
Vivo
Samples

15. Handheld mass spectrometry system:
15

16. 16

17. Handheld mass spectrometry system:
■ The system contains three
primary components:
– (i) Syringe pump to deliver a
defined water volume (4 to 10
ml);
– (ii) Small diameter (800 mm)
polytetrafluoroethylene
(PTFE) tubing conduits;
– (iii) Handheld pen-sized probe
for direct sampling of
biological tissues.
17

18. MasSpec Pen Design and Operation
18
The probe tip is designed with three ports:
1. an incoming port that delivers a single
water droplet to the probe tip
(conduit 1)
2. a central port for gas delivery
(conduit 2)
3. an outgoing port to transport molecular
constituents in the water droplet from
the tissue to the mass spectrometer
(conduit 3)
T=0s T=2s T=3s
The main component of the handheld pen-
sized probe is a 3D printed
polydimethylsiloxane (PDMS) tip

19. How Effective is this Handheld Pen?
19
What Basically is detected
in the sample by this pen?

20. Metabolites Detected by MasSpec Pen in a Cell
20
Source: LIPID MAPS Lipidomics Gateway Human Metabolome Database
215.031
327.23
906.64
600.513
885.55
790
834.5
773.5
145.05
146.04
174.04
175.02
215.02
327.3
906.63
600.52
885.55
790
834.5
773.4
145.04
146.05
174.04
175.01
Hexose
FA
ST
CER
PI
PE
PS
PG
Glutamine
Glutamate
N-acetylaspartic acid
Ascorbic acid
Identification of molecular ions
Theoretical
m/z
Measured
m/z

21. 21
Molecular Analysis of Thin Tissue Sections
using the Mass Spec Pen
MasSpec Pen analysis of a mouse brain tissue section and a background region of glass slide (no
sample) are shown (average of n=5 mass spectra each)

22. How Effective is this Handheld Pen?
22
How does it detects normal cells and
cancerous cells
in the sample?

23. 23
MolecularAnalysis ofThinTissue
Sections using the MasSpec Pen

24. Molecular analysis of thin tissue sections
using the MasSpec Pen
24

25. Abundance of cell metabolites in
normal and cancerous tissue
25
100
70
50
2
30
5
20
80
100
10
10
50
40
20
Iodine
FA
Ascorbic acid
CER
PI
PE
Glutamine
Identification of molecular ions
Cancerous tissue Normal tissue

26. How Effective is this Handheld Pen?
26
What makes this Pen Nondestructive in
Nature?

27. Nondestructive molecular analysis of
human tissue samples
27
Images show a lung adenocarcinoma
tissue sample before, during, and
after the MasSpec Pen analysis.
The automated and time-controlled
operational steps of the MasSpec Pen
prevent tissue damage because the
tissue is only exposed to the small
water droplet and not to the vacuum
used to transport the droplet from the
reservoir to the mass spectrometer

28. Comparison Between the Masspec Pen
Mass Spectra Obtained From the Frozen
Tissue Section and Fresh Tissue Piece
28

29. 29
Comparison between the MasSpec Pen
mass spectra obtained from the tissue
section and fresh tissue piece.

30. 30
20
20
70
50
90
20
30
90
70
100
ST
CER
PI
PE
PS
Identification of molecular ions
Fresh Tissue Frozen Tissue
Cell metabolites obtained from the tissue
section and fresh tissue piece.

31. Patient demographics of the 253 human
tissue samples used in this study
31

32. Intrasample analysis of histologically
distinct and cancer margin tissue regions
32

33. 33
Intrasample analysis of histologically
distinct and cancer margin tissue regions

34. 34
Comparison between mass spectra obtained
from the regions of tissue section
30
20
10
5
5
40
70
50
40
30
20
100
100
60
50
Ascorbic acid
FA
PI
PS
PE
RelativeAbundance
Identification of molecular ions
Region 3 Region 2 Region 1

35. How effective is this handheld pen?
35
How is MasSpec pen used for In Vivo cancer
detection ?

36. In vivo analysis of a mouse model of
human breast cancer during surgery
36

37. In vivo analysis of a mouse model of
human breast cancer during surgery
37

38. 38
In vivo and Ex vivo comparison of a
mouse model of human breast cancer
during surgery

39. Questions yet to be Answered
■ Can this pen be used for detection other cancers?
■ When it will be available for actual cancer
detection in hospitals?
39

40. References
1. Diagnosis of Cancer by Metabolic Signatures using Mass Spectrometric
Imaging (October 2014)
2. Beger, R., 2013. A review of applications of metabolomics in
cancer. Metabolites, 3(3), pp.552-574.
3. J. Maygarden, F. C. Detterbeck, W. K. Funkhouser, Bronchial margins in lung
cancer resection specimens: Utility of frozen section and gross evaluation. Mod.
Pathol. 17, 1080–1086 (2004)
4. Lu, M., Faull, K.F., Whitelegge, J.P., He, J., Shen, D., Saxton, R.E. and Chang,
H.R., 2007. Proteomics and mass spectrometry for cancer biomarker
discovery. Biomarker insights, 2, p.117727190700200005.
5. A. M. Nick, R. L. Coleman, P. T. Ramirez, A. K. Sood, A framework for a
personalized surgical approach to ovarian cancer. Nat. Rev. Clin. Oncol. 12,
239–245 (2015)
40

41. THANK YOU…
41

42. Effect of MasSpec Pen contact time on the mass spectra obtained
42