Aug 26, 2022
PSI FluorCAM protocol for Rapid Light Curve with far red pre-ilumination.
- 1University of Technology Sydney
Protocol Citation: Andrei Herdean 2022. PSI FluorCAM protocol for Rapid Light Curve with far red pre-ilumination.. protocols.io https://dx.doi.org/10.17504/protocols.io.x54v9deypg3e/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 26, 2022
Last Modified: August 26, 2022
Protocol Integer ID: 69222
Abstract
PSI FluorCAM protocol for Rapid Light Curve with far red pre-ilumination.
PSI FluorCAM protocol for Rapid Light Curve with far red pre-ilumination.
PSI FluorCAM protocol for Rapid Light Curve with far red pre-ilumination.
Load the following script in to the FluorCam software
TS=50ms
;Rapid light curve in Actinic light 2 for TOMI-2
;version November 11, 2020
;with FilterWheel
;high-resolution CCD - TOMI-2
;estimated time: app. 372 s
;for successful measurement check Light curve settings manual
ADD1=0
Act2=0
Act1=0
;
include default.inc ;Includes standard options, do not remove it !
include light.inc ;Includes standard options, do not remove it !
include FW.inc ;Includes standard options, do not remove it !
Shutter=2
Sensitivity=30.6
Super=60
ADD2=10
__LightA=24.122
__LightB=-61.463
;3 = 10.903
;4 = 35.025
;5 = 59.147
;6 = 83.269
;7 = 107.391
;8 = 131.513
;9 = 155.635
;10 = 179.757
;15 = 300.367
;20 = 420.977
;25 = 541.587
;30 = 662.197
;35 = 782.807
;40 = 903.417
;These are the light intensities used for the Rapid light curve
I1 = 3
I2 = 3
I3 = 3
I4 = 3
I5 = 3
I6 = 3
I7 = 3
I8 = 3
I9 = 3
I10 = 3
I11 = 4
I12 = 5
I13 = 6
I14 = 7
I15 = 8
I16 = 9
I17 = 10
I18 = 12
I19 = 17
I20 = 20
I21 = 25
I22 = 30
I23 = 35
I24 = 40
I25 = 45
__LightIntensity=
;_____________________________________________________________________________________________________________
;There is a pre-illumination phase with only far red light, followed by determination of Fo and Fm
<0s>=>SET_FILTER(CHL)
Preillumination=60s
<0s>=>add2(Preillumination)
start = Preillumination;
;-------------------------------------------------------------------------------------------------------------
;*** Fo Measurement ******************************************************************************************
;-------------------------------------------------------------------------------------------------------------
F0duration = 5s
F0measure = 1s
start + <0,F0measure..F0duration>=>mfmsub
start + <0s>=>checkPoint,"startFo"
start + =>checkPoint,"endFo"
;
;-------------------------------------------------------------------------------------------------------------
;*** Saturating Pulse & Fm Measurement ***********************************************************************
;-------------------------------------------------------------------------------------------------------------
PulseDuration = 800ms;
Fmstart = start + F0duration+mfmsub_length
;
=>SI_Act2(I25)
=>SatPulse(PulseDuration)
=>act2(PulseDuration)
Fmstart+<200ms, 200ms + mfmsub_length .. PulseDuration-mfmsub_length>=>mfmsub
;
=>checkPoint,"startFm"
=>checkPoint,"endFm"
=>checkPoint,"timeVisual"
;
;*** Actinic light Exposure *******************************
Pause = 2*TS
ALduration = 30s
ALmeasure = ALduration/6
;Protocol contains 5 minutes of illumination with low light in order to keep photosynthesis active while samples are brogth to target temperatures by the thermocycler
ALstart1 = Fmstart + PulseDuration + Pause
ALstart2 = ALstart1 + ALduration + Pause
ALstart3 = ALstart2 + ALduration + Pause
ALstart4 = ALstart3 + ALduration + Pause
ALstart5 = ALstart4 + ALduration + Pause
ALstart6 = ALstart5 + ALduration + Pause
ALstart7 = ALstart6 + ALduration + Pause
ALstart8 = ALstart7 + ALduration + Pause
ALstart9 = ALstart8 + ALduration + Pause
ALstart10 = ALstart9 + ALduration + Pause
;this is where the Light Curve starts
ALstart11 = ALstart10 + ALduration + Pause
ALstart12 = ALstart11 + ALduration + Pause
ALstart13 = ALstart12 + ALduration + Pause
ALstart14 = ALstart13 + ALduration + Pause
ALstart15 = ALstart14 + ALduration + Pause
ALstart16 = ALstart15 + ALduration + Pause
ALstart17 = ALstart16 + ALduration + Pause
ALstart18 = ALstart17 + ALduration + Pause
ALstart19 = ALstart18 + ALduration + Pause
ALstart20 = ALstart19 + ALduration + Pause
ALstart21 = ALstart20 + ALduration + Pause
ALstart22 = ALstart21 + ALduration + Pause
ALstart23 = ALstart22 + ALduration + Pause
ALstart24 = ALstart23 + ALduration + Pause
ALstart25 = ALstart24 + ALduration + Pause
;
=>SI_Act2(I1)
=>SI_Act2(I2)
=>SI_Act2(I3)
=>SI_Act2(I4)
=>SI_Act2(I5)
=>SI_Act2(I6)
=>SI_Act2(I7)
=>SI_Act2(I8)
=>SI_Act2(I9)
=>SI_Act2(I10)
=>SI_Act2(I11)
=>SI_Act2(I12)
=>SI_Act2(I13)
=>SI_Act2(I14)
=>SI_Act2(I15)
=>SI_Act2(I16)
=>SI_Act2(I17)
=>SI_Act2(I18)
=>SI_Act2(I19)
=>SI_Act2(I20)
=>SI_Act2(I21)
=>SI_Act2(I22)
=>SI_Act2(I23)
=>SI_Act2(I24)
=>SI_Act2(I25)
;
ALstart =
ALstart => act2(ALduration)
;
;******* Kautsky Effect Measurement **********************************
;
;ALstart + TS => mfmsub
ALstart#=>mfmsub
;
;******************* Ft'& Fm' definition **********************************
PulseLStart = ALduration - PulseDuration - mfmsub_length
;
FtL = ALstart + PulseLStart - PulseDuration
FtL#=>mfmsub
;
PulseL = ALstart + PulseLStart
PulseL - TS=> SI_Act2(I25)
PulseL=>SatPulse(PulseDuration)
PulseL#<200ms, 200ms + mfmsub_length .. PulseDuration-mfmsub_length>=>mfmsub
;
Last = ALstart25+ALduration+2*mfmsub_length
Last + <0s, 1s .. 5s>=>mfmsub
;--------------------------------------------------------------
;
=>checkPoint,"startFm_Lss1"
=>checkPoint,"endFm_Lss1"
;
=>checkPoint,"startFm_Lss2"
=>checkPoint,"endFm_Lss2"
;
=>checkPoint,"startFm_Lss3"
=>checkPoint,"endFm_Lss3"
;
=>checkPoint,"startFm_Lss4"
=>checkPoint,"endFm_Lss4"
;
=>checkPoint,"startFm_Lss5"
=>checkPoint,"endFm_Lss5"
;
=>checkPoint,"startFm_Lss6"
=>checkPoint,"endFm_Lss6"
;
=>checkPoint,"startFm_Lss7"
=>checkPoint,"endFm_Lss7"
;
=>checkPoint,"startFm_Lss8"
=>checkPoint,"endFm_Lss8"
;
=>checkPoint,"startFm_Lss9"
=>checkPoint,"endFm_Lss9"
;
=>checkPoint,"startFm_Lss10"
=>checkPoint,"endFm_Lss10"
;
=>checkPoint,"startFm_Lss11"
=>checkPoint,"endFm_Lss11"
;
=>checkPoint,"startFm_Lss12"
=>checkPoint,"endFm_Lss12"
;
=>checkPoint,"startFm_Lss13"
=>checkPoint,"endFm_Lss13"
;
=>checkPoint,"startFm_Lss14"
=>checkPoint,"endFm_Lss14"
;
=>checkPoint,"startFm_Lss15"
=>checkPoint,"endFm_Lss15"
;
=>checkPoint,"startFm_Lss16"
=>checkPoint,"endFm_Lss16"
;
=>checkPoint,"startFm_Lss17"
=>checkPoint,"endFm_Lss17"
;
=>checkPoint,"startFm_Lss18"
=>checkPoint,"endFm_Lss18"
;
=>checkPoint,"startFm_Lss19"
=>checkPoint,"endFm_Lss19"
;
=>checkPoint,"startFm_Lss20"
=>checkPoint,"endFm_Lss20"
;
=>checkPoint,"startFm_Lss21"
=>checkPoint,"endFm_Lss21"
;
=>checkPoint,"startFm_Lss22"
=>checkPoint,"endFm_Lss22"
;
=>checkPoint,"startFm_Lss23"
=>checkPoint,"endFm_Lss23"
;
=>checkPoint,"startFm_Lss24"
=>checkPoint,"endFm_Lss24"
;
=>checkPoint,"startFm_Lss25"
=>checkPoint,"endFm_Lss25"
;
;******************* Ft' definition **********************************
;
=>checkPoint,"startFt_Lss1"
=>checkPoint,"endFt_Lss1"
;
=>checkPoint,"startFt_Lss2"
=>checkPoint,"endFt_Lss2"
;
=>checkPoint,"startFt_Lss3"
=>checkPoint,"endFt_Lss3"
;
=>checkPoint,"startFt_Lss4"
=>checkPoint,"endFt_Lss4"
;
=>checkPoint,"startFt_Lss5"
=>checkPoint,"endFt_Lss5"
;
=>checkPoint,"startFt_Lss6"
=>checkPoint,"endFt_Lss6"
;
=>checkPoint,"startFt_Lss7"
=>checkPoint,"endFt_Lss7"
;
=>checkPoint,"startFt_Lss8"
=>checkPoint,"endFt_Lss8"
;
=>checkPoint,"startFt_Lss9"
=>checkPoint,"endFt_Lss9"
;
=>checkPoint,"startFt_Lss10"
=>checkPoint,"endFt_Lss10"
;
=>checkPoint,"startFt_Lss11"
=>checkPoint,"endFt_Lss11"
;
=>checkPoint,"startFt_Lss12"
=>checkPoint,"endFt_Lss12"
;
=>checkPoint,"startFt_Lss13"
=>checkPoint,"endFt_Lss13"
;
=>checkPoint,"startFt_Lss14"
=>checkPoint,"endFt_Lss14"
;
=>checkPoint,"startFt_Lss15"
=>checkPoint,"endFt_Lss15"
;
=>checkPoint,"startFt_Lss16"
=>checkPoint,"endFt_Lss16"
;
=>checkPoint,"startFt_Lss17"
=>checkPoint,"endFt_Lss17"
;
=>checkPoint,"startFt_Lss18"
=>checkPoint,"endFt_Lss18"
;
=>checkPoint,"startFt_Lss19"
=>checkPoint,"endFt_Lss19"
;
=>checkPoint,"startFt_Lss20"
=>checkPoint,"endFt_Lss20"
;
=>checkPoint,"startFt_Lss21"
=>checkPoint,"endFt_Lss21"
;
=>checkPoint,"startFt_Lss22"
=>checkPoint,"endFt_Lss22"
;
=>checkPoint,"startFt_Lss23"
=>checkPoint,"endFt_Lss23"
;
=>checkPoint,"startFt_Lss24"
=>checkPoint,"endFt_Lss24"
;
=>checkPoint,"startFt_Lss25"
=>checkPoint,"endFt_Lss25"
;
;
;END ******************************************************************