| 
| 
|
|---|---|---|
TRANSCRIPT
Present were: 22
orchidnut (Sam - Nebraska)
nodosa (Ed - San Antonio TX)
marylois (Lois - NW Louisiana)
sparkysteve (Steve - Boca Raton FL
Jane5536 (Jane - L.I. NY)
Ellen,Smithtown,NewYork
ScottMcPhee (location unknown)
soobie me (Sue - NC)
KB Barrett (Kathy - northern CA)
Josh319 (Josh - IA)
JCY8S (John - Arcadia CA)
BCPRESS (Bert - Miami FL)
peeteilis (Tom - Algonquin Island NY)
johngingarland (John - Garland TX)
paulav (Paula - Boca Raton FL)
gaillevy (Gail - Boca Raton FL)
onelaelia (Ursula - Caracas, Venezuela)
AORCHID (Art - Simpsonville SC)
knudy (Kip - MI)
Fleur (Tasmania, Australia)
jchandler (Josh - IA)
Evlyn (northern Louisiana)
nodosa (Ed in SAT)
Hi, all. Can't believe I got here early. Must be a GREAT program on for
tonite.
orchidnut (Sam in Lincoln, NE)
I heard you have some pix tonight, Ed
nodosa (Ed in SAT)
Sam, I got to tell you: Safari gets a centerfold this evening. Gonna make
Internet history.
orchidnut (Sam in Lincoln, NE)
Cool! hehehehe
nodosa (Ed in SAT)
Step one of flask case design is to select a comfortable chair and work
surface. Something that really fits. You'll spend long hours in flasking.
Once a suitable work area is selected, we will try to duplicate it in the
flask case setup.
Let's be sure we understand what a flask case is: it is a sterile work area
that is kept that way by air from a positive pressure plenum (box for holding
air) that is in turn supplied from a negative pressure plenum. The two may
be stacked over, under or side by side to meet the space available.
Ambient air (air from the work area) is drawn into the fan which is the
heart of the negative plenum. There it is compressed and blown thru a very
fine filter, emerging as sterile air in the work area. That's all there
is to a flask case. Questions up to this point?
When the air stream is established in a flasking case, we insert two filters
in it. A common furnace filter is used to prefilter incoming air and a fine
filter is used to provide sterile air.
JCY8S (John in Arcadia, CA)
Ed - what is a ''fine'' filter? How fine?
nodosa (Ed in SAT)
Fine filters are of two types: HEPA filters and DOP filters. HEPA filters
are High Energy Particle Arresting filters that are 97 - 99 per cent effective.
DOP filters are hospital grade filters that are 95 -99 per cent effective.
DOP filters are about a third less expensive than HEPA, so we use DOP. The
extra per cent or two isn't worth the difference.
Fleur
Is this the same principle as a laminer flow cabinet Ed?
nodosa (Ed in SAT)
Both DOP and HEPA filters are available from W. W. Grainger, Inc. which
has outlets in most major cities.
soobie me (Sue in NC)
How often does the filter have to be replaced?
nodosa (Ed in SAT)
Recap: we now have a comfortable place to work, an understanding of the
principle of sterile air and a brief overview of filters and sources of
supply. Are we all ready now to assemble these things to serve our flasking
needs?
soobie me (Sue in NC)
I guess it depends on how dirty the air is! ready to go on!!!
nodosa (Ed in SAT)
Filters are replaced when we begin to get contaminated flasks. I don't know
how long that is - we do lots of work and our flasks are not contaminating
after 5+ years of use.
nodosa (Ed in SAT)
For many reasons, we chose to make a floor mounted case and put it on rollers.
We move it out to use it and back to store it. NOW YOU GET A PIX of our
case with its dust cover on (an old bed sheet). [picture]
nodosa (Ed in SAT)
Well, OK, you get a side view. Note at the top right, we have a piece of
acrylic partly over the work area. This reduces contaminatin enormously.
[picture]
Next should be a front view with covers over the filters. [picture]
JCY8S (John in Arcadia, CA)
Ed- the filters are on the bottom?
nodosa (Ed in SAT)
The tan covers inside the work area are close fitting to protect the filters
when they are not in use. The section nearest the floor is the intake filter
and the section inside the work area is the outlet. Again, the acrylic shield
can be seen. This is not normally installed on a laminar flow work station,
but it can save a lot of contamination - especially if the operator has
a cold!
The white 'box' at the top is a fluorescent light. It is a 2 tube fixture
made to slip into a suspended ceiling. We reversed the lens and inverted
the fixture. Makes a dandy work light and is outside the sterile area. PIX--same as
above
Now let's see if we can take the covers off the filters. [picture]
orchidnut (Sam in Lincoln, NE)
Does the cover effect the airflow e.g. turbulance?
nodosa (Ed in SAT)
You'll note the bottom (intake) filter is a common home furnace filter:
Home Depot for a buck. The top filter is the fine filter, which is deeply
pleated to provide more filtering area. This makes a depth of about 12",
so you have to allow for that in your design. Leave filter depth and room
for the air to move around the pleats.
Sam, if you mean the clear acrylic, yes it has an effect. It channels the
air into your lap and threatens to freeze never never land.
johngingarland
*G*
nodosa (Ed in SAT)
While we are here, notice we extended the bottom supports to make the cabinet
more stable. The base is 2X4 lumber, the cabinet is weather-resistant 3/8"
plywood.
Let's turn the cabinet around. [picture]
Sparing no expense, we used a regular extension cord for a power cord. We
store it
coiled on small boat deck cleats at opposing corners. Note the back is one
piece and is installed with screw nails. When we need to maintain fine filter
or squirrel cage fan, we just take the back off.
We'll get the back off and look at the equipment layout. [picture]
Now you can see those massive pleats in the fine filter and see why the plenum must be rather large to provide room for them plus the air.
onelaelia (Caracas)
How long have you had it? Don't the metal parts rust?
nodosa (Ed in SAT)
The squirrel cage fan was salvaged from an old central heat unit. It is
mounted to the underside of a heavy piece that goes all the way out front
to make the work area.
Fleur (Tasmania)
Is that foam you have used to seal the openings?
nodosa (Ed in SAT)
This unit has been in use about five years. No reason for the metal parts
to rust.
nodosa (Ed in SAT)
Good eye, Fleur. Yeah, we went to a lot of trouble to seal everything with
urethane foam weather stripping. Waste of time. It won't hurt anything if
it leaks a little, though that is no excuse for sloppy work. Only place
it mustn't leak is around the fine filter. All air entering the work area
must pass thru that.
KB Barrett (Kathy in N Calif)
That's a really impressive set up Ed. I was anticipating finding out how
dissimilar flasking would be from home canning... not anything like what
you've got going!!!
nodosa (Ed in SAT)
The cabinet really stands up straight. The list is due to poor camera work.
We put a coat of primer and two coats of latex semi-gloss enamel on all
wood surfaces. That way, they can be washed (although we never have scrubbed
them).
Remember, the basics we're seeing can be applied to a table top unit. There, you might want to put the fan on top. One thing for sure, there is no such thing as an easily portable flasking case that will do a good job year after year.
sparkysteve
Ed--has anyone ever tried to use UV fluorescent lights, the type they use
at restaurants --or isn't it necessary?
nodosa (Ed in SAT)
Years ago, UV lights were in all sterile work stations. I can't imagine
the eye damage that was done. Today, OSHA would have your lunch for using
one. There is no way to fully protect your eyes from damage if you install
a UV sterilizer. DON'T DO IT.
OK, let's look again at the work area. [picture]
orchidnut (Sam in Lincoln, NE)
Can you get fine filters that are flat to save space, and not pleated like
yours?
nodosa (Ed in SAT)
Note that heavy metal frame on the fine filter. That's handy - you can mount
it just by supporting it IF you seal around the fine filter so air doesn't
leak and bypass it.
Sam, you must use the pleated filters. Static pressure (air resistance) is enormous in these filters because they are so fine. The huge area permits the volume of air past the filter. A flat one would blow very little air and burn out the fan.
sparkysteve
Can I ask a really dumb question Ed? Which way does the air flow? From the
blue filter up?
nodosa (Ed in SAT)
The work surface and walls look pretty impressive. I could tell you they
are custom stainless steel. Truth is, they are 20' wide aluminum flashing.
You cut and fit it with a large pair of shears, then glue it down with contact
cement. A little silicone calk in the seams and corners and you have your
custom top.
Air comes in thru the blue filter, is compressed and forced up by the squirrel cage, comes thru the back of the fine filter and emreges to flood the work area with sterile air.
If this covers the case, we'll look at the product. Any questions before we move on?
Fleur (Tasmania)
Looks impressive though. Just one... do you bleach the inside before you
start a new batch, and if so does it have any effect on the aluminum.
nodosa (Ed in SAT)
When we first do flasks, they go into an incubator. The red rubber mat barely
visible on the bottom is a heat mat that keeps the minumum temp at 75 deg.
F. We used to give the flasks supplemental growth lamp light but now we
grow them at very low light values. [picture]
Yes, Fleur, we use clorox water in the work area and yes, it causes pitting
in the aluminum. When it gets bad enough, we'll just put another layer on
top of the base layer. Should be able to use 3 or 4 layers before we have
to rework.
The incubator is under the main the greenhouse. It is just framed in, then
covered with foil faced styrofoam(the kind they put under vinyl siding.
In the winter, we have plexiglas doors that install all the way across to
keep the flasks snug.
Fleur (Tasmania)
What is the light source?
nodosa (Ed in SAT)
In stage 2, we move flasks to a growing cabinet in the open greenhouse.
Supplemental light is installed, but we rarely use it except on really cold
nights when we use it for a bit of extra heat. In that case, we cover the
frong with a sheet of plastic film. [picture]
The light source is just stray daylight, Fleur. Very new flasks want very
little light. You want them to be sugar feeders so they will develop roots
and differentiate. Too much light will stimulate chlorophyll production
and make the little plants try to be air feeders - and there's no CO2 source
in a flask.
Fleur (Tasmania)
What temp is the mat set for?
nodosa (Ed in SAT)
Excuse me - cover the front with sheet plastic. Again, we have insulated
a light frame with insul board.
orchidnut (Sam in Lincoln, NE)
What type of jars do you use?
nodosa (Ed in SAT)
The mat in the incubator is set for 70 deg F. We do not use a mat in the
growing cabinet. The bottles are 500 and 1,000 ml square French bottles.
As you can see, stacking is important to us. Also, you work French bottles
from the side, reducing your exposure to contamination. Finally, they cost
less than Erlenmeyer flasks.
nodosa (Ed in SAT)
Before we finish, I would like for you to meet our lab technicians. Good
teks are hard to find and we always give kudos to ours because she is one
of the very best. [picture]
Fleur (Tasmania)
The flasks most commonly used here are the 'Plastic' type. with a vent in
the top. Do yours have the vent?
nodosa (Ed in SAT)
That about does it for me. Are there questions or comments. We have no secrets,
so ask what you like.
orchidnut (Sam in Lincoln, NE)
What size of fan do you have in your cabinet?
Fleur (Tasmania)
No wonder you have such good flasks with a helper like that. (laughing my
head off here)
sparkysteve (
I remember her! I think I met her in a bar one night!
nodosa (Ed in SAT)
We use standard one hole rubber stoppers, Fleur. Size 8.5 for the 500's
and size 10 for the 1,000's. We pack the hole moderately firm with non-absorbent
cotton. We have tried many plastic flasks and always come back to the French
bottles. I know many who see it the other way, however.
sparkysteve
Ed--do you break them to replate?
nodosa (Ed in SAT)
That's where we found her, Steve. She didn't mention you as a reference.
Sam, I have no idea what size our fan is - whatever came out of the old
furnace. I should thing you would need to move 100 - 400 cfm,just as a guess.
Fleur (Tasmania)
I had one batch of flasks that became contaminated soon after I got them
home. All with the 'hole in the lid' I have always blamed the hole for the
contamination. Some flasks do not have this feature and I have not had any
problems with them.
nodosa (Ed in SAT)
We try not to break flasks to replate or to plant out. On replates, we empty
flasks by using a long pair of tweezers. A hardware store near us sells
them for 5 bucks each to unjam garbage disposals. When we want to plant
out, we fill the flask with tepid water, let it stand 5 minutes or so, then
ease the contents out with another pair of tweezers.
orchidnut (Sam in Lincoln, NE)
Do you have a reostat installed to control the fan speed?
nodosa (Ed in SAT)
Fleur, any time you move a flask you run a risk of contamination. Spores
can lurk for a long time, then sprout when jarred loose.
You could have a rheostat, Sam, or just wire it thru a light dimmer if the wattage isn't too high. No use complicating it, though. Ours works just fine.
Fleur
Why do the flasks have this vent?
nodosa (Ed in SAT)
One point on contamination. Watch flasks carefully. At the first sign of
contamination, try to burn it out by focusing sunlight on it thru a large
magnifying glass. If that doesn't do it, try to replate the flask, leaving
an ample area around the contamination undisturbed.
johngingarland
Ed, what fungicide do you reccomend for planting out?
KB Barrett (Kathy in N Calif)
Fleur, I *think* it's for gas exchange, but I really know nothing about
flasking. I think Andy uses plastic wrap to cover his flask openings, which
amazed me since I didn't think anything went thru saran!
nodosa (Ed in SAT)
Several reasons for the vent, Fleur. First, when you autoclave air must
vent out of the flask and the pressure vessel before the pressure is allowed
to rise. Air will compress; steam is a solid. Next there is gas exchange
in the flask. A fully sealed flask would rapidly kill the plants thru oxygen
poisoning.
Fleur
I know it sounds like a silly question but I'm going to ask it anyway...
Would the UV from a sunbed kill contamination?
nodosa (Ed in SAT)
We do not use a fungicide for planting out, John. The little plants have
enough to cope with just coming into our world. If you have damping off
problems, put a half inch layer of milled sphagnum on top of the pot medium
and plant thru that.
Fleur
Thank you Ed, I thought it was something like that. Maybe the packing got
moist at some stage allowing entry of bugs.
KB Barrett (Kathy in N Calif)
Fleur, I'll bet one of those laser pens would give better 'spot' decontamination.
Don't laugh, there have been studies that show they improve wound healing.
Go figure!
nodosa (Ed in SAT)
Fleur - yes, and the plants and your eyes if you don't protect them.
marylois (NW Louisiana)
I have goggles to use with my UVB machine.
nodosa (Ed in SAT)
That might be a good thought, Kathy. I have a pretty strong laser pen -
I'll try it. Only problem might be the thickness of the glass. Re the packing
for the vent: this is not to make them air tight. The purpose is to construct
an infinite baffle thru which gasses can exchange while pathogens 'get lost'.
nodosa (Ed in SAT)
Marylois, did you ever try to fit goggles on an aurantiaca seedling?
Fleur
Me too Lois. Although my machine is sitting in the garage at the moment
It has to be dismantled to get it upstairs*sigh*
KB Barrett (Kathy in N Calif)
Ed, if it works make sure you call it the 'Barrett Technique of Decontamination'
or some such! He he! I gotta get famous somehow!
nodosa (Ed in SAT)
Last call on the topic, then let's release all to informal chat. Thanks
for your attention!
marylois (NW Louisiana)
Ah, I thought human eyes were the concern.
nodosa (Ed in SAT)
Your name will be up in laser lights, KB.
KB Barrett (Kathy in N Calif)
Ed, with my luck I'll become an Urban Legend, right up there with the microwaved
poodle!!
Fleur
One thought.. Going back to Andy and his plastic film over the vents. Basic
laws of keeping things sterile.. Keep it dry.. keep it dry.. keep it dry.
As long as the packing material does not get moist bugs will not pass through.
Plastic film may keep it dry.
Fleur
Ed, thank you so very much, this was a very good topic to have. Next one...
how to pollinate a flower you can't even see?
nodosa (Ed in SAT)
Fleur, we put 4.5 X 4.5 inch aluminum squares (cut from household foil)
over our stoppers before autoclaving. If a cork pops you can usually reseat
it if you do so before handling the flask much. The aluminum covers keep
the flasks clean indefinitely. We prepare lots of flasks, then store them
for use when needed. Storage life seems indefinite.
johngingarland
Thanks, Ed! Most Enjoyable! Also enjoy your column in SWROGA!
Fleur
Ah, the ones I received had no covering over the stoppers/vents at all just
a cotton wick.
marylois (NW Louisiana):
Tolja it would be inimitable!
KB Barrett (Kathy in N Calif)
Ed, is there a time of year that you sit down to flask? or just as pods
ripen and and on an as needed basis?
WE FLASK WHEN PODS RIPEN. ONE OF THE MOST VALUABLE THINGS A HYBRIDIZER CAN DEVELOP IS A SYSTEM FOR TELLING WHEN TO FLASK A SEED POD. OURS WORKS WELL FOR US. WE ALSO TRY TO SET TWO PODS FOR EVERY CROSS IN CASE WE LOSE ONE. WE TEND TO LET REPLATE WORK STACK UP UNTIL WE HAVE TIME TO DO IT (NOT A RECOMMENDED PRACTICE) AND THEN DO SEVERAL DAYS OF REPLATING. IF WE SEE A FLASK THAT IS PROTOCORMING OR JUST NOT DOING WELL, WE'LL REPLATE IT ON SHORT NOTICE, SOMETIMES CHANGING THE MEDIUM.
nodosa (Ed in SAT)
Many thanks to Lois, who did all the pix work. Great job despite a bit of
operator trouble at first. Thanks!
johngingarland
Or is that, 'Columns?' [Indeed it is columnS - Ed is a major supporter of
the SWROGA News!...mlg]
KB Barrett (Kathy in N Calif)
I can just see Ed flasking by the light of the first full moon of spring'
cause the Farmers Almanac says that's the most auspicious time. Ah, another
urban legend is born!
DON'T LAUGH, KATHY. WE TRY TO POLLINATE USING THE HAWAIIAN MOON CALENDAR. CAN'T SAY IT WORKS BUT WE NEED ALL THE HELP WE CAN GET.
Fleur
One last question Ed, does the size of flask you use reflect the size of
plant you are expecting.
KB Barrett (Kathy in N Calif)
Good question fleur! Or the viability of the cross? If lots of offspring
are expected or only a few?
Lanceps (Thamina from Manhattan Beach, Calif.)
nodosa, I've bought flasks with foil over the stoppers.
nodosa (Ed in SAT)
Fleur, as a rule, we do mother flasks (seed sowing) in 500 ml's. Depending
on the plantlet size, the first replate will be 500 or 1,000. Phals are
usually quite large and need 1,000's. We believe in replating often, so
we don't put a whole lot of medium in each flask. Keeps the roots manageable.
nodosa (Ed in SAT)
Foil is quick, cheap and effective, Thamina, so now you know our criteria!
sparkysteve (of Boca Raton, Florida)
Ed--what do you have to say about a flask sent through airline shipment--any
comments?
Fleur
Thank you Ed, I would only need small flasks then for replating masdevallias.
KB Barrett (Kathy in N Calif)
Fleur, going to start hybridizing your award winners?
marylois (NW Louisiana)
That would be the real kick!
KB Barrett (Kathy in N Calif)
Of course that's predicated on your ability to see the flower...
marylois (NW Louisiana)
...that's the kicker. *S*
nodosa (Ed in SAT)
We don't have much trouble shipping flasks, Steve. Years ago, customs insisted
on autoclaving flasks and infusing methyl bromide. We just heat sealed the
flasks in plastic tube and kept shipping.
Lanceps (Thamina from Manhattan Beach, Calif.)
The mother flask contains the seed, standard size. the small plants are
replated to less crowded flasks to finish growing--- at least thats the
way my flasker does it . I tell him how many flasks I want replated from
the mother flask,2 or 4.
Lanceps (Thamina from Manhattan Beach, Calif.)
My flasker uses canning jars now,punches a hole in the lid,uses cotton and
tape. Easier to pot out from the wide mouth.
Fleur
I would love to Kathy, but the basic polinating is still very hard for me.
Some of the plants are so small I have trouble finding the bits.
marylois (NW Louisiana)
Can you lock one up in a sealed room with a proper insect, Fleur. :-)
Fleur
Lois, I wish. *grin* send me the ecuadorian orchid fly and I will give it
a go. I have the use of a lamineer flow cabinet any time I want it but...
marylois (NW Louisiana)
Let's all give Ed a big hand for filling in so beautifully on VERY short
notice. Many thanks, Edward.
nodosa (Ed in SAT)
Fleur, I would try to decide on one size of flask - two at the most. Flasks
get expensive by the 100's. An oversize flask won't hurt the plants. You
certainly could standardize on 250 ml Erlenmeyers or 250 ml French squares.
Both are readily avialable and inexpensive. Find a dairy that switched from
glass to cartons and you'll probably find a batch of 250 ml bottles that
were used for heavy cream. Most take a size 7 stopper and work well.
Fleur
Thank you, Thank you. Ed, what a wonderful idea, the local dairy, I would
never have thought of them.
KB Barrett (Kathy in N Calif)
Yes, Ed. It was a wonderful presentation and done on such short notice!!!
sparkysteve (of Boca Raton, Florida)
The pictures are excellent
nodosa (Ed in SAT)
We'll try to archive ASAP. Right now we have a major problem dealing with
a pending USDA decision to permit import of potted orchid plants (Phalaenopsis).
Could spell dire times for many US growers.
Evlyn
From what country, Ed?
KB Barrett (Kathy in N Calif)
Meaning what, Ed? Right now they only can bring in flasks, and they want
to bring in grown plants?
nodosa (Ed in SAT)
The import proposal refers to Taiwan but if implemented it would cover all
qualifying nations. Our concern is the homegrown, quality pot plant Phal
will be supplanted by an import that does not hold up on the shelf or in
the customer's use. That could kill the entire orchid pot plant industry
that many of us have struggled to create.
nodosa (Ed in SAT)
I must leave, too. Thanks to all for a fine evening. I hope to see you again
real soon.
sparkysteve (of Boca Raton, Florida)
you did great ED---thanks
KB Barrett (Kathy in N Calif)
Looks like its time to hit the lights, Sparky!
|
|
|
|
|---|---|---|