The Virtual Tour

BIOLUMINESCENT IMAGING OF STREPTOCOCCUS PYOGENES IN NASOPHARYNGEAL INFECTIONFaraz Alam1, Colin Bateman1, Claire Turner1, Ken Smith2, Siouxsie Wiles1,3 & Shiranee Sriskandan1

1 Dept. of Infectious Diseases & Immunity, Imperial College London, London W12 0NN, UK. 2Dept. of Pathology & Infectious Diseases, Royal Veterinary College, Hertfordshire AL9 7TA , UK.

3 Dept. of Molecular Medicine & Pathology, University of Auckland, 1142, New Zealand.

Current Models

Live Mouse

Current Models

Live Mouse

Cull

Organs

Current Models

Live Mouse

Cull

Organs

Plate

Bacteria

Current Models

Live Mouse

Cull

Organs

Plate

Bacteria

Current Models

Single Time Point

Bact

eria

/ g

ram

of ti

ssue

Time

Live Mouse

Cull

Organs

Plate

Bacteria

Direct Culture

Direct Culture

0 7 14 210

20

40

60

80

100

Males Females

Time (days)

Per

cen

t C

arri

age

Infection Longevity

Inflammation in the Nasopharynx

Inflammation in the Nasopharynx

Turner & Sriskandan, 2007

Impact of covRS on carriage

0 5 10 15 200

20

40

60

80

100

Wild type

covRS Mutant

Days Post Inoculation

Per

cen

t C

arri

age

Impact of covRS on carriage

Airborne Transmission

Airborne Transmission

hrs

Dir

ect

Nas

al S

amp

le c

ou

nts

(cf

u)

in R

ecip

ien

t M

ice

0 20 40 60 801

10

100

1000

10000 2 seed mice per cage

3 seed mice per cage

4 seed mice per cage

Airborne Transmission

Photorhabdus luminescens

Bioluminescence Imaging

Light

Camera

Bioluminescence Imaging

Light

Camera

Bioluminescence Imaging

Bioluminescence Imaging

Camera

Light

Growth Kinetics

Growth Kinetics

Time

Opt

ical

Den

sity

Luminescence(R

LU)

0 2 4 6 8 100.0

0.5

1.0

0

5000

10000

15000

20000

Wild type

Spy0535 Mutant

Bioluminescent strain

Luminescent signal

In vivo Growth Kinetics

In vivo Growth Kinetics

cfu

/ n

ose

100

101

102

103

104

105

106

107

108

109Wild type vs Luminescent

Day 1 Day 1 Day 1Day 3 Day 3 Day 3

Wild type vs Knockout Luminescent vs Knockout

In vivo Growth Kinetics

cfu

/ n

ose

100

101

102

103

104

105

106

107

108

109Wild type vs Luminescent

Day 1 Day 1 Day 1Day 3 Day 3 Day 3

Wild type vs Control Luminescent vs Control

In vivo Growth Kinetics

cfu

/ n

ose

100

101

102

103

104

105

106

107

108

109Wild type vs Luminescent

Day 1 Day 1 Day 1Day 3 Day 3 Day 3

Wild type vs Control Luminescent vs Control

In vivo Growth Kinetics

Days Post Inoculation

Fit

nes

s re

lati

ve t

o w

ildty

pe

0 1 2 3 40

1

2

3

4

Luminescent Strain

Control strain

100 101 102 103 104 105 106 107 10810000

100000

1000000

Total CFU

To

tal F

lux (

ph

oto

ns/

seco

nd

)

Click # FMA2010062408431524 Jun 2010 8:43:15Em filter=OpenBin:M (8), FOV10, f1, 5mCamera: 13195, EEV

User: FAGroup: H494 Day 2Experiment: Animal Number: G, BLView: Dorsal

70000

60000

50000

40000

30000

20000

ImageMin = -4984.5Max = 79669p/sec/cm^2/sr

Color BarMin = 20000Max = 72038

bkg subflat-fieldedcosmic

ROI 1

Bioluminescence in the nasopharynx

FA20100813120726_SEQ

FA20100120123803

4

5

6

7

8

910

4

2

3

4

5

6

7

8

910

5

Color BarMin = 4000

Max = 1e+05

FA20100120124351 FA20100120125227

FA20100120125833 FA20100120130548 FA20100120131155

Longitudinal imaging

Longitudinal imaging

Vaccination Study

PBS

Vacc

inat

edH

eat K

illed

Bac

teria

Day 1 Day 35Day 14 Day 28

Vaccination StudyPB

S Va

ccin

ated

Hea

t Kill

ed B

acte

ria

Day 1 Day 2 Day 3 Day 4

Vaccination Study

Time Course

Days

Lu

min

esce

nce

(p

ho

ton

s/se

con

d)

0 1 2 3 4 510000

100000

1000000

Heat Killed H347PBS

Cfu

/ G

ram

Heat Killed H347 PBS100

101

102

103

104

105

106

107

108

Mucosal Transmission

Summary

Summary

Bact

eria

/ g

ram

of ti

ssue

Time

Single Time Point

Photograph/ Nasal Sample

Summary