Fluorescence Imaging Site B

1
Fluorescence Imaging Site B

• Life in the Atacama 2005Science Technology
  WorkshopJanuary 6-7, 2005
• Shmuel Weinstein
• Kim Warren-Rhodes
• Greg Fisher

2
Introduction

• This presentation is a review of the data
  products produced by the Fluorescence Imager and
  the fluorescent probes during the Science Ops of
  Site B, 2004 field season in the Atacama Desert
• Outline
• Overview of FI capabilities
• FI results on a locale by locale basis
• Summary

3
Overview of 2004 FI capabilities

• View
• 10 cm x 10 cm directly under belly of Zoe,
• 180 ?m resolution
• Illumination/Excitation
• 1 J/flash Xenon flashlamp
• Four fiber bundle illumination channels
• Two intensities of white light, UV(325 nm 375
  nm), blue
• (425 nm 475 nm), green(530 nm 560 nm), and
  red(643 nm 688 nm)
• Detection
• R, G, B, chlorophyll(670 nm 810 nm), DNA probe
• (485 nm 535 nm), Protein probe(590 nm 650
  nm), two other emission channels(435 nm 485
  nm) and (800 nm 850 nm)

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The Dyes Used in the Field
5
The Operation

• Sequence
• PRGB, chlorophyll (blue excitation), water
  spray, BRGB, chlorophyll, DNA, Protein,
  fluorescent probes spray, DDNA, Protein
• Later added chlorophyll check (green excitation)
  after every chlorophyll
• File Nomenclature
• Z040917_1531_013FIP54X62MS_rawS

Date
Time
FI
Sequence Step
Filter Set
Image Process
Image Quality
P, B, or D
rawS
45X74M Chlor 54X74M ChlorChk 45X51M
DNA 54X62M Protein ND1RGB RGB comp
N no flash F flash S F-N subtracted
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Rating System

• Four-point rating system to be unambiguous
• Biological morphology
• Chlorophyll positive
• DNA dye positive
• Protein dye positive

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What did we find?

• Sol 2, Locale 1

The Positive Effect of Water on the Sample
8
What did we find?

• Sol 2, Locale 1

Before and After DNA Dye
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What did we find?

• Sol 2, Locale 1

Before and After Protein Dye
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What did we find?

• Sol 2, Locale 1

Rating 3 out of 4 (morphology, chlorophyll, DNA)
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What did we find?

• Sol 2, Locale 2

Rating 2 out of 4 (morphology, chlorophyll)
12
What did we find?

• Sol 3, Locale 5

Rover movement (wind, motors?) created blurry dye
images and did not allow proper dye analysis. Are
these mini lichens or endolithic life, or did the
water wash off dust of mineral fluorescence? Ratin
g 1 out of 4 (chlorophyll)
13
What did we find?

• Sol 3, Locale 6

Chlorophyll positive in a number of small
regions. Dim, but real increases in both dye
channels (quantitative comparisons on the .tif
images) Rating 3 out of 4 (no definitive
morphology)
14
What did we find?

• Sol 5, Locale 19

Slight fluorescence in both before and after dye
in DNA and Protein- but NO increase due to
dye. Rating 2 out of 4 (morphology, chlorophyll
with chlorophyll check) or is it greater?
15
What did we find?

• Sol 6, Locale 19

chlorophyll chlorophyll check DNA
Protein
A hole filling the field-of-view with a single
rock at the bottom in the center. Sign of life?
-) In the small region of interest from the
full field-of-view (above), note the chlorophyll
signature in center mineral fluorescence in
lower right Rating 1 out of 4
16
Conclusion

• In the best estimation, the FI detected both
  chlorophyll and dyes, although there was not a
  conclusive, unambiguous detection of life in any
  one sample, using all four criteria established
  for the science ops.
• Better dye penetration of lichens would improve
  that.
• Wind and/or other vibrations and rover movement
  will likely prevent image subtraction as a way of
  on-board image processing.