scope.property_changer("SOLA","White_Enable",1)
## basic settings
run_number=3
interval= 300 # set to 15 minutes (in seconds)
timepoints= 864
## less changed settings
save_dir=r"E:\Folks\Nate\20160425_%s" % run_number
pos_record=r"E:\Folks\Nate\20160425_%s\PositionHistory.csv" % run_number
#### Getting stage locations
positions_file=u'C:/Users/Administrator/Desktop/MicroscopeControl/MicroscopeControl/HalfChipPointsList.pos'
positions=StagePositions.positionList_parser(open(positions_file).read())
order=sorted(positions.keys(),key= lambda x: int(x[3:]))
### running everything
#setting up path
if os.path.exists(save_dir) is False: os.mkdir(save_dir)
fout=open(pos_record,"w")
header="%s;%s;%s;%s;%s;%s;%s;%s;%s;%s;%s;%s;%s;%s\n" % ("Pos",
"Time",
"x_set",
"y_set",
"z_set",
"afc_set",
def multi_point_timelapse(
scope, save_dir, positions_file, illumination_pattern_object, n_timepoints, frequency, file_name_prefix="Timelapse"
):
"""
args:
scope -- initalized scope object -- see Microscopes.Microscope
"""
#####################################################
### Pre Imaging ###
#####################################################
#### Getting stage locations
positions = StagePositions.positionList_parser(open(positions_file).read())
pos_order = sorted(positions.keys(), key=lambda x: int(x[3:])) # numerical order
#### Getting the channels that will be taken
# channels=illumination_pattern_object.channels
# channels=["BF","qGFP","qmCherry"]
channels = ["qDIC", "qGFP", "qCherry"]
#### Set up the to save images in a structured set of directories
if os.path.exists(save_dir) is False:
os.mkdir(save_dir)
for pos_label in pos_order:
pos_dir = os.path.join(save_dir, pos_label)
if os.path.exists(pos_dir) is False:
os.mkdir(pos_dir)
for channel_name in channels:
channel_dir = os.path.join(pos_dir, channel_name)
if os.path.exists(channel_dir) is False:
os.mkdir(channel_dir)
#### Set up the log file
pos_stats_file_path = os.path.join(save_dir, "Position_Stats.tsv")
fout = open(pos_stats_file_path, "w")
pos_record_values = [
"Pos",
"Time",
"x_set",
"y_set",
"z_set",
"afc_set",
"x_1",
"y_1",
"z_1",
"afc_1",
"x_2",
"y_2",
"z_2",
"afc_2",
"channels",
]
header = "\t".join(pos_record_values) + "\n"
fout.write(header)
fout.close()
#####################################################
### Imaging Loop ###
#####################################################
try:
### interate over timepoints
for t in range(0, n_timepoints):
print ["init", "init", "move", "foc ", "imag", "save", "stats"]
acquisition_start_time = time.time()
for pos_label in pos_order:
t1 = time.time()
## lookup where to go
pos = positions[pos_label]
x = pos.get_x()
y = pos.get_y()
z = pos.get_z()
afc = pos.get_afc_offset()
print "Locations settings: %s\t%s x\t%s y\t%s z\t%s offset" % (pos_label, x, y, z, afc)
## move scope to x,y
t2 = time.time()
scope.move_XYstage(x, y)
## see where the scope is
x1, y1, z1, afc1 = scope.get_stage_loc()
## focus the scope using afc offset
t3 = time.time()
scope.move_to_offset(afc, post_focus_delay=0.1, attempt_delay=0.1)
## see where the scope is now
x2, y2, z2, afc2 = scope.get_stage_loc()
## figure out the channels to image
# c_to_image=conditional_illuminations.get_channels(pos_label,t)
# c_stack_settings=conditional_illuminations.get_stack_settings(pos_label,t)
c_to_image = [Illuminations.BF2, Illuminations.dimGREEN, Illuminations.dimRED]
# c_to_image=["qDIC","qGFP","qCherry"]
c_stack_settings = {"qDIC": [5, 1]}
## take the images
t4 = time.time()
images, metadata = scope.multichannel_zstack(c_to_image, z_params=c_stack_settings, timed=True)
## save the images
t5 = time.time()
for c in range(0, len(c_to_image)):
stack = images[c]
md_stack = metadata[c]
try:
channel_name = c_to_image[c].name
except:
channel_name = c_to_image[c]
full_save_dir = os.path.join(save_dir, pos_label, channel_name)
full_file_name = "%s_%s_c%s%s_t%s" % (file_name_prefix, pos_label, c, channel_name, t)
FileOut.stack_writer(full_save_dir, full_file_name, stack, md_stack[len(md_stack) / 2])
t6 = time.time()
###log some stats
channels_aqcuired_string = "" # this is in progress
# for c in c_to_image:
# name=c.name
# intensity=c.SOLA_intensity
# length=c.exposure
# if name in c_stack_settings:
# z=c_stack_settings[name][0]/c_stack_settings[name][1]+1
# else: z=1
# channel_string=""
pos_record_stats = (
pos_label,
t,
x,
y,
z,
afc,
x1,
y1,
z1,
afc1,
x2,
y2,
z2,
afc2,
channels_aqcuired_string,
)
pos_record_stats = [str(i) for i in pos_record_stats]
pos_record_stats = "\t".join(pos_record_stats)
pos_record_stats += "\n"
fout = open(pos_stats_file_path, "a")
fout.write(pos_record_stats)
fout.close()
t7 = time.time()
times = [t1, t2, t3, t4, t5, t6, t7]
times = ["%.2f" % (i - times[0]) for i in times]
times_lab = ["Start", "Find Where to Go", "Move Stage", "Focused", "Imaged", "Saved", "Saved_Stats"]
for i in range(0, len(times)):
print times[i], times_lab[i]
##return the to first location
scope.move_to_offset(afc)
xnext = positions[pos_order[0]].get_x()
ynext = positions[pos_order[0]].get_y()
scope.move_XYstage(xnext, ynext)
scope.mmc.enableContinuousFocus(False)
### wait for the start of the next acquisition
acquisition_finish_time = time.time()
until_next = frequency - (acquisition_finish_time - acquisition_start_time)
verbose_sleeper(until_next)
print "Timelapse completed"
results = "Scan Completed"
except:
print "UH OH!"
tb = StringIO.StringIO()
exc_type, exc_value, exc_traceback = sys.exc_info()
traceback.print_exception(exc_type, exc_value, exc_traceback, file=tb)
results = (
"There was some sort of error, "
"your scan most likely did not finish."
"The error was \n\n %s" % tb.getvalue()
)
return results
### know what you are doing! ###
### ###
##########################################################
scan_visibility=False
import PlateScan
import EmailFromNTHAYER
import StringIO
import StagePositions
import traceback
import sys
pos_string=open(pos_file).read()
positions=StagePositions.positionList_parser(pos_string)
try:
scope=scopeConfig.DGLab_Leica2()
scope.mmc.setProperty("XYStage","Speed",str(stageSpeed))
except:
tb=StringIO.StringIO()
exc_type, exc_value, exc_traceback = sys.exc_info()
print "Could not initialize scope. Try restarting python and/or scope"
traceback.print_exception(exc_type,
exc_value,
exc_traceback,
file=tb)
print tb.getvalue()
raise
try:
scan_results=PlateScan.plateScan(scope,
def legacy():
"""
This is the code that I had originall wrote to do this.
This code was really messy, but worked, so I am leaving it here.
It has been rewritten in the above functions, but could be useful to look
at.
"""
## initialize the scope
scope=DGLab_Leica2.DGLab_Leica2()
## set up the
rows=string.ascii_uppercase[1:7]
columns=range(2,12)
sites=range(0,225)
pos_string=open("PositionList_96well_15x15sites.pos").read()
positions=StagePositions.positionList_parser(pos_string)
scope.mmc.enableContinuousFocus(True)
scope.set_illumination(Illuminations.Cy5)
#start_time=time.asctime().replace(" ","_").replace(":","_")
start_time="Thu_Aug_13_14_38_51_2015"
done_list={}
well_counter={}
## if this is a continuation of an old scan, this will add on to the
## existing log file.
if os.path.exists("Scan_%s.txt" % start_time):
completed_fin=open("Scan_%s.txt" % start_time,"r")
for line in completed_fin:
site=line.split(";")[0]
try:
if "No cells of interest" not in line.split(";")[1]:
well=site.split("-")
if well not in well_counter:
well_counter[well]=0
well_counter+=1
except:
pass
completed_fin.close()
status_out=open("Scan_%s.txt" % start_time,"r")
status_string=status_out.read()
status_out.close()
status_out=open("Scan_%s.txt" % start_time,"w")
status_out.write(status_string)
else:
status_out=open("Scan_%s.txt" % start_time,"w")
data_dir="Scan_%s" % start_time
if os.path.isdir(data_dir) is False:
os.mkdir(data_dir)
order=-1
for row in rows:
## flip the order that the columns are gone through
order=order*-1
for column in columns[::order]:
well="%s%s" % (row,column)
if well not in well_counter:well_counter[well]=0
if well_counter[well]<15:
## reset the well count for this new well.
well_count=well_counter[well]
## iterate through the site in this well.
for site in sites: ## Test case
if "%s%s-Site_%s" % (row,column,site) not in done_list.keys():
done_list["%s%s-Site_%s" % (row,column,site)]=True
print ">Looking at %s%s-Site_%s" % (row,column,site)
x,y=positions["%s%s-Site_%s" % (row,column,site)].get_xy()
## take an image to use for a trigger
scope.set_illumination(Illuminations.qCy5)
scope.mmc.setXYPosition("XYStage",x,y)
scope.mmc.snapImage()
img=scope.mmc.getImage()
## show image
plt.imshow(img)
plt.draw()
## analyze this image
cois=Threshold.coi_finder(img,
threshold=1000,
min_area=100,
max_area=1000)
if cois:
do_stack=True
progress_line=">%s%s-Site_%s;cois=%s\n" % (row,column,site,cois)
else:
do_stack=False
progress_line=">%s%s-Site_%s;No cells of interest\n" % (row,column,site)
status_out.write(progress_line)
## run the stack if there was a COI found
if do_stack:
print ">Stack at %s%s-Site_%s" % (row,column,site)
well_count+=1
well_counter[well]+=1
channels=[Illuminations.DIC,
Illuminations.GFP,
Illuminations.qCy5,
Illuminations.DAPI]
scope.mmc.enableContinuousFocus(False)
mda,md=scope.multi_channel_acquisition(channels)
FileOut.mda_save(out_dir=data_dir,
prefix="%s%s-Site_%s" % (row,column,site),
mda=mda,
md=md,
channels=channels)
print "Stack taken at %s%s-Site_%s" % (row,column,site)
## turn AFC back on
scope.mmc.enableContinuousFocus(True)
## move onto the next well if we have acquired enough
if well_count > 15:
break
## break out of the well if you can't focus
if scope.mmc.isContinuousFocusLocked() is False:
time.sleep(5)
if scope.mmc.isContinuousFocusLocked() is False:
print "Couldnt focus 3"
break
## break out of the column if you still cant focus
if scope.mmc.isContinuousFocusLocked() is False:
time.sleep(5)
if scope.mmc.isContinuousFocusLocked() is False:
print "Couldnt focus 2"
break
## break out of the row if you STILL cant focus.
if scope.mmc.isContinuousFocusLocked() is False:
time.sleep(5)
if scope.mmc.isContinuousFocusLocked() is False:
print "Couldnt focus 1"
break
status_out.close()
