CS CSI: I: DUND DUNDEE EE Th The e Fo Fore rensic nsic To - - PowerPoint PPT Presentation

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CS CSI: I: DUND DUNDEE EE Th The e Fo Fore rensic nsic To - - PowerPoint PPT Presentation

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CS CSI: I: DUND DUNDEE EE Th The e Fo Fore rensic nsic To Tool olkit kit Meet the Team Meet the Team What is Forensic Science? What is Forensic Science? Why Forensic Science? Why Forensic Science? Why Forensic Science? Why

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CS CSI: I: DUND DUNDEE EE

Th The e Fo Fore rensic nsic To Tool

  • lkit

kit

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SLIDE 2

Meet the Team

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SLIDE 3

Meet the Team

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SLIDE 4

What is Forensic Science?

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SLIDE 5

What is Forensic Science?

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SLIDE 6

Why Forensic Science?

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SLIDE 7

Why Forensic Science?

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SLIDE 8

Why Forensic Science?

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SLIDE 9

Why Forensic Science?

“There is Forensics and then there is Science.”

Sue Black – OBE, FRSE, Director of CAHID and Professor of Anatomy and Forensic Anthropology at the University of Dundee

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SLIDE 10

Public blic Perception: rception: The e CSI Effect fect

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SLIDE 11

Public blic Perception: rception: The e CSI Effect fect

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Public blic Perception: rception: The e CSI Effect fect

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SLIDE 13

Why Forensic Science?

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The Experts

Home Office Centre for Applied Science and Technology (CAST) Chemist

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Meeting the Experts

NAME: Prof. Niamh Nic Daeid TITLE: Professor

  • f Forensic

Science NOTE: Over 150 published papers

  • n forensics
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Meeting the Experts

NAME: Kenny Laing TITLE: Fingerprint Enhancement Specialist for Scottish Police Service Authority

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Meeting the Experts

NAME: Dr. Lucina Hackman TITLE: Forensic Anthropologist QUOTE: “ I can dismember a body in 20mins and still have time for a cup of tea!”

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The Solution: The Forensic Toolkit

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SLIDE 19

FluID: Body Fluid Detector CELL FREE ALL IN ONE

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How would it work?

NASAL SAL MUCUS CUS SALIVA LIVA BLOOD OOD SEMEN MEN LACTOF CTOFERR RRIN HAEMOG EMOGLOB OBIN N A AND D HAEMOG EMOGLOB OBIN N B SPERMI ERMIDIN INE ODORAN ORANT BINDIN NDING PROTEI OTEIN

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How does it work?

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How does it work?

Blood Saliva Nasal Mucus Semen

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Where did we get our parts?

PLASMID

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General Idea OV OVEREXPRES EREXPRESS PU PURIFY RIFY CH CHARACTERI ARACTERIZE ZE

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SLIDE 25

Blood Detection

Haptoglobin ptoglobin Haemoglobin emoglobin B Haemoglobin emoglobin A

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SLIDE 26

Haptoglobin

Successfully purified human an hapto ptoglo globin bin from E.coli!

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Haemoglobin B

A-HIs

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Blood Detection

Haptoglobin ptoglobin Haemoglobin emoglobin B Haemoglobin emoglobin A Next xt steps: ps: Perform chem emic ical al cross ss linki nking ng assay says s with hapto ptoglobin globin and haemoglobin moglobin B

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SLIDE 29

Semen Detection

Targe rget: t: Sperm ermidi idine ne

PotD

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SLIDE 30

Semen Detection

8 10 12 14

  • 500

500 1000 1500

Volume (ml) UV Absorbance (mAU)

Size Exclusion Chromatogram of PotD

A12 B12

Successfully purified PotD tD from

  • E. coli!
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SLIDE 31

PotD/Spermidine Binding

Measured Intrinsic tryptophan fluorescence

Excitation= 280nm Emission = 350nm

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SLIDE 32

How do we detect?

  • Sy

Synt nthe hetic tic

  • Fl

Fluo uore rescen scent t la late tex x beads beads

  • Visible

Visible

  • La

Labe bel l fr free ee

  • On

One pro e probl blem: em: they are always fluorescing!

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SLIDE 33

Future Work

  • Continue characterisation of

binding protein/ligand interactions

  • Ensure attachment of fluorescent

bead does not interfere with these interactions

  • Test for any potential interactions

between the four binding proteins

  • Design a suitable container and

select an appropriate medium

  • Make sure device does not

damage or degrade DNA!

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SLIDE 34

Mathematical Modeling

R₀ =

𝐽𝑜𝑗𝑢𝑗𝑏𝑚 𝑑𝑝𝑜𝑑𝑓𝑜𝑢𝑠𝑏𝑢𝑗𝑝𝑜 𝑝𝑔 𝑞𝑝𝑢𝐸 𝐽𝑜𝑗𝑢𝑗𝑏𝑚 𝑑𝑝𝑜𝑑𝑓𝑜𝑢𝑠𝑏𝑢𝑗𝑝𝑜 𝑝𝑔 𝑡𝑞𝑓𝑠𝑛𝑗𝑒𝑗𝑜𝑓

Semen men: : PotD tD and Spermidine rmidine Bindi nding ng Model el

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SLIDE 35

Ageing of Fingerprints

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Principal Component Analysis: PCA

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Composite Contribution

*Data ata court urtesy esy of Dr.

  • Dr. Frick,

ck, Curtin tin Unive iversi rsity ty Australia tralia

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SLIDE 38

Squalene Epoxide

Over 7 days Under 7 days

Squalene Squalene Epoxide Lanosterol Lanosterol Synthase

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General Process

OV OVEREXPRES EREXPRESS PU PURIFY RIFY CH CHARACTERI ARACTERIZE ZE

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So far…

Lanosterol synthase has been successfully overexpressed

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LS Inactivation

Active ive sites es

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Next Steps

  • Continue characterization of

lanosterol synthase in native state

  • Mutate lanosterol synthase active

site residues

  • Ensure this does not affect binding

with squalene epoxide

  • Attach fluorescent bead and

characterize binding parameters

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SLIDE 43

Mathematical Modeling

V₀ =

𝐽𝑜𝑗𝑢𝑗𝑏𝑚 𝑑𝑝𝑜𝑑𝑓𝑜𝑢𝑠𝑏𝑢𝑗𝑝𝑜 𝑝𝑔 𝑚𝑏𝑜𝑝𝑡𝑢𝑓𝑠𝑝𝑚 𝑡𝑧𝑜𝑢ℎ𝑏𝑡𝑓 𝐽𝑜𝑗𝑢𝑗𝑏𝑚 𝑑𝑝𝑜𝑑𝑓𝑜𝑢𝑠𝑏𝑢𝑗𝑝𝑜 𝑝𝑔 𝑡𝑟𝑣𝑏𝑚𝑓𝑜𝑓 𝑓𝑞𝑝𝑦𝑗𝑒𝑓

Squalene ualene epoxid

  • xide

e and d lanosterol nosterol synthase nthase bindin nding g model del

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Chromate Biosensor

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Chromate Biosensor

BBa_K1 a_K1058 58008 08

Beij ijin ing g Inst stit itute ute of Tech chno nolo logy y (2013) 13)

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Chromate Biosensor

GFP ChrB

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In theory…

ChrB GFP +Cr(VI) +Cr(VI)

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Testing our System

Probl

  • blem:

em: GFP expression detected in the absen sence ce of chromate.

  • mate.
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Chromate Modeling

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Bone Incision Experiments

Sliding force

W = kPS

Wear volume Force Wear constant

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Conclusion

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Conclusion

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Acknowledgements

  • Tracy

cy Palmer mer

  • Frank

nk Sargen gent

  • Fordyce

dyce Davidson dson

  • Fatima

ima Ulhuq uq

  • Lucas

as J. M. Moya

  • Sue Black

ck

  • Niamh

mh Nic Nic Daeid Daeid

  • Kenny

ny Laing ng

  • Grant

nt Buchan hanan an

  • Marta

ta Albare areda da

  • Lucia

ia Licand andro ro Lado Lado

  • Gary

y Callon lon

  • James

es Moir

  • Avril

il Smart rt

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SLIDE 54