def elisa_wash(protocol: protocol_api.ProtocolContext,
p300m: InstrumentContext,
reservoir: [Labware],
plate: Labware,
num_washes: int,
wash_volume: int):
"""
TODO: consider splitting into a load function and a remove function? this one is large. Is that modularity useful?
# Maybe modularity is useful for multiple plates? Rather than changing this function to accommodate multiple plates
# TODO: modularity here is almost certainly useful. Can reuse funcs for FACs prep
Args:
protocol: From robot
p300m: P300 multichannel
reservoir: A 12 well trough containing ELISA wash buffer
plate: The plate to be washed
num_washes: number of plate washes to perform
wash_volume: volume of buffer to wash each well with
Returns:
nothing
"""
# dispenses per aspiration to minimize movement and counter initialization
dispenses_per_load = int(300/wash_volume)
# Calculate extra volume
well_counter = 0
vol_counter = 0
# Pick up tips
# TODO: adjust num presses and increment
p300m.pick_up_tip(p300m.tip_racks[0].well('A1'), presses=2, increment=.05)
for i1 in range(num_washes):
# reset aspiration height
index = 0
# TODO: add disposal volume?
"""
~~~~~~~~~~~~~~~~~~
Handle adding wash to plate. Calculate max number of dispenses per source load.
Loop and increment index appropriately. This calculation would be automatic,
but volume removed from source well has to be tracked manually to prevent empty.
~~~~~~~~~~~~~~~~~~
"""
for i2 in range(int(12/(dispenses_per_load))):
# Establish a source well. well_counter should increment before well runs out of liquid
# src must be established within this loop
if vol_counter >= 9.7:
# If most of the volume from the trough is gone, move to next well and reset volume count
well_counter += 1
vol_counter = 0
src = reservoir[0].columns()[well_counter]
wells = plate.rows()[0][index: index+dispenses_per_load]
# TODO: figure out volume count problem...works but not soon enough. well_counter iterates after two washes
# TODO: maybe tips don't go down to bottom of well and that's why?
p300m.well_bottom_clearance.aspirate = 5
p300m.well_bottom_clearance.dispense = 10
p300m.distribute(volume=wash_volume, source=src, dest=wells,
new_tip='never', disposal_volume=0, blow_out=True)
# Increment index by number of dispenses per aspiration
index += dispenses_per_load
# Count volume consumed from current trough well
vol_counter += (.3 * 4)
# Tried distribute and keep getting tip already attached or tip not attached error
"""
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Mix and dispense most of the liquid into the trash. Manual flicking still necessary.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
"""
# protocol.delay(seconds=30)
# # Set clearance for tip for aspiration
# p300m.well_bottom_clearance.aspirate = 0.1
# # Mix and transfer volume from plate to trash.
# p300m.transfer(volume=wash_volume, source=plate.rows()[0],
# dest=p300m.trash_container.wells(0), disposal_volume=wash_volume,
# new_tip='never',) #mix_before=(2, (wash_volume*.75)), aspirate_speed=5, dispense_speed=12,
p300m.home()
protocol.pause(msg="{} washes done! Resume washing by clicking the 'Resume' button!".format(i1+1))
protocol.comment("{} washes complete! You may proceed with your protocol.".format(num_washes))
p300m.return_tip()
p300m.home()
def elisa_wash(protocol: protocol_api.ProtocolContext,
p300m: InstrumentContext, reservoir: [Labware], plate: Labware,
wells_to_fill: [[]], num_washes: int, wash_volume: int):
"""
TODO: consider splitting into a load function and a remove function? this one is large. Is that modularity useful?
# Maybe modularity is useful for multiple plates? Rather than changing this function to accommodate multiple plates
# TODO: modularity here is almost certainly useful. Can reuse funcs for FACs prep
Args:
protocol: From robot
p300m: P300 multichannel
reservoir: A 12 well trough containing ELISA wash buffer
plate: The plate to be washed
num_washes: number of plate washes to perform
wash_volume: volume of buffer to wash each well with
Returns:
nothing
"""
# dispenses per aspiration to minimize movement and counter initialization
dispenses_per_load = int(300 / wash_volume)
# Calculate extra volume
well_counter = 0
vol_counter = 0
# Pick up tips
# TODO: adjust num presses and increment
p300m.pick_up_tip(p300m.tip_racks[0].well('A1'), presses=3, increment=.05)
for i1 in range(num_washes):
# reset aspiration height
p300m.well_bottom_clearance.aspirate = 1
index = 0
# TODO: add disposal volume?
"""
~~~~~~~~~~~~~~~~~~
Handle adding wash to plate. Calculate max number of dispenses per source load.
Loop and increment index appropriately. This calculation would be automatic,
but volume removed from source well has to be tracked manually to prevent empty.
~~~~~~~~~~~~~~~~~~
"""
# for i2 in range(int(12/(dispenses_per_load))):
# Establish a source well. well_counter should increment before well runs out of liquid
# src must be established within this loop
src = reservoir[0].columns()[well_counter]
# location is a list of pairs defining locations of wells
location = wells_to_fill
print("location{}".format(location))
# subset to locations in the distribution range (number of dispenses per aspiration)
locs = location[index:index + 3]
print(locs)
# for all lists in locs, take the second (1) index aka the row
# rows = plate.wells(*locs[2])
# if multiple rows entered into parentheses, each row returned will be a list
# so plate.row(1) yields [[wells B1-B12]] and plate.rows(1,2) yields [[wells B1-12], [C1-12]]
"""
The logic below should underlie transforming csv files created by the plate mapper
into a format to access plate wells
"""
rows = []
cols = []
for i in location:
cols.append(i[0])
if i[1] not in rows:
rows.append(i[1])
plate_map = plate.rows(*rows)
updated = []
for i in plate_map:
# there shouldnt be a +1 for normal list slicing but apparently slicing is noninclusive or something
updated.append(i[min(cols):max(cols) + 1])
print(updated)
# TODO: figure out volume count problem...works but not soon enough. well_counter iterates after two washes
# TODO: maybe tips don't go down to bottom of well and that's why?
for i in cols:
p300m.distribute(volume=wash_volume,
source=src,
dest=rows[i],
new_tip='never',
disposal_volume=0,
blow_out=True)
# Increment index by number of dispenses per aspiration
index += dispenses_per_load
# Count volume consumed from current trough well
vol_counter += (.3 * 8)
if vol_counter >= 9.7:
# If most of the volume from the trough is gone, move to next well and reset volume count
well_counter += 1
vol_counter = 0
# Tried distribute and keep getting tip already attached or tip not attached error
"""
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Mix and dispense most of the liquid into the trash. Manual flicking still necessary.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
"""
# Set clearance for tip for aspiration
p300m.well_bottom_clearance.aspirate = 0.1
# Mix and transfer volume from plate to trash.
p300m.transfer(
volume=wash_volume,
source=plate.rows()[0],
dest=p300m.trash_container.wells(0),
disposal_volume=100,
new_tip='never',
mix_before=(2, 80),
aspirate_speed=5,
dispense_speed=12,
)
p300m.home()
protocol.pause(
msg="{} washes done! Resume washing by clicking the 'Resume' button!".
format(i1 + 1))
protocol.comment(
"{} washes complete! You may proceed with your protocol.".format(
num_washes))
p300m.return_tip()
p300m.home()