Ian,

Here are our programs. The Brain blocks have been fixed for several
weeks prior to processing, hence the short processor fixation time. You
will probably need to vary the dehydration and clearing times for your
tissues:

The Leica TP1020 is a carousel type tissue processor with agitation,
vacuum and fume extraction.

Routine Processing Programs

Program			1	2	3	4	5
Station Solution		Routine-Overnight,Two-Basket-Routine,
Urgent, Routine-Weekend,Brain
1	Formalin		1:00	1:15	0:10	1:30 (No Vacuum)
10 min
2	Formalin		1:00	1:15	0:20	1:30	10 min
3	70% Ethanol		0:30	1:15	0:05	1:00	1:00
4	Absolute Ethanol	1:30	1:15	0:10	2:00	4:00
5	Absolute Ethanol	1:30	1:15	0:10	2:00	4:00
6	Absolute Ethanol	2:15	1:15	0:10	2:00	4:00
7	Xylene		0:30	1:15	0:10	1:30	2:00
8	Xylene		1:00	1:15	0:10	1:30	2:00
9	Xylene		1:30	1:15	0:10	1:30	2:00
10	Wax	1:00		1:15	0:05	1:30	2:00
11	Wax	1:00		1:15	0:10	1:30	3:00
12	Wax	2:00		1:15	0:10	2:00	3:00
Total Time			14:57	15:12	2:17	19:42	27:20

Immediate Program Start
1.	Press  (Program displayed always last one used)
2.	Choose Program using <+> or <-> Keys (see above)
3.	Press  to begin program

Urgent Program Start

1.	For programs less than 8 hours duration a warning code will
appear
2.	Check that the wax in the paraffin stations is molten, and if
so:
3.	Press  and  simultaneously to override warning code
and commence processing
 

Delayed Start (Weekend)

This program does not use vacuum until Step 2.

To finish at 7am Monday Morning:

1.	Press   
2.	Select Program 4 (using <+> or <-> Keys)
3.	Press  Key
4.	Enter 2 as days of delay (<+> or <-> Keys)
5.	Press  Key for hours
6.	Enter "11" (<+> or <-> Keys) - start at 11am Sunday Morning
7.	Press   to commence program

To check if programmed starting time leads to an acceptable end of run
time press 

Security
To protect program settings from accidental changes, the key functions
of the control panel can be locked.

1.	Press  and hold for 5 seconds
2.	"LOCKED" will be displayed

To unlock, Press  and hold for 5 seconds.

Pausing a Program to add more cassettes

1.	Press 
2.	Press  to raise basket (If vacuum is on, this might
take some time)
3.	Add cassettes
4.	Press  to return basket to station
5.	To resume process press 

Stop/Abort a Program

1.	Press 
2.	When "STOP?" message appears, to abort program, press 
again
 

End of Process

1.	"DONE" is displayed.
2.	Alarm sound is heard every 30 seconds
3.	Press any key to stop alarm

Maintenance

Please refer to Maintenance Manual and Instrument Instruction Manual. 
The solutions need to be changed after every five runs by the person on
call.
Daily Maintenance

1.	Check liquid level in each station
2.	Check quality of each fluid
3.	Clean rims of all stations and seal of each lid
4.	Wipe control panel clean

After Five Runs

1.	Discard first formalin, rotate and add fresh formalin to last
formalin station (Station 2)
2.	Discard 70% ethanol and add fresh 70% ethanol
3.	Discard first absolute ethanol, rotate and add fresh absolute
ethanol to last absolute ethanol station (Station 6)
4.	Discard first xylene, rotate and add fresh xylene to last xylene
station (Station 9)

After Ten Runs

1.	As per above maintenance plus
2.	Discard first wax
3.	Rotate wax stations
4.	Add fresh molten wax to last wax station (station 12)

Monthly

1.	Lift carousel cover to upper end position
2.	Clean carousel axle with a clean cloth
3.	Apply a thin coat of machine oil

If the machines are turned off for a period long enough to solidify the
wax, it is important to place a rod, or something similar, to break the
vacuum seals of the wax lids, otherwise the container can warp.


Recycling

*	Xylenes can be recycled. 
*	Alcohols to be collected and recycled, unless they contain
xylene, then discard (ie alcohols after xylene in dewaxing and alcohol
wash)

When Processing Goes Wrong 

Tissue can appear so poorly processed, that the blocks smell of xylene
or the clearing agent used in processing. In these cases, blocks can be
melted and returned to fresh molten wax overnight to improve
infiltration before embedding.

If tissue appears soft at the embedding stage, then dehydration is often
the problem. This is common if tissues are too thick, resulting in poor
movement of processing fluids around and into the tissue. Another sign
is the presence of indentations from the cassette and lid on the tissue.
These tissues require reprocessing. 

While the tissue is still hot (ie the wax is still molten) blot the
tissue dry, place it back in its cassette and place the cassette in 10%
formalin for reprocessing. This procedure dramatically improve the
results in at least 90% of cases. The excellent results are probably due
to the protective nature of the wax present in the adequately processed
portions of the block. This insulates the tissue from the harmful
effects of ethanol on the adequately processed portions of the tissue
preventing the tissue from becoming hard and brittle (Johnson 2003).

Johnson (2003) Histologic 36(1):21-22.

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318




-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ian
Montgomery
Sent: Wednesday, 1 November 2006 8:48 PM
To: Histonet
Subject: [Histonet] Leica TP1020


    After years hand processing I've finally got myself a Leica TP1020.
But, I'm having a few teething problems and with the work piling up the
problems are weighing heavily on my shoulders. Processing schedules, any
hints and tips? I've Googled and searched the Histonet archive but
without much success. The two major projects I have at the moment are
salmon muscle and mouse brain. I've tried several schedules but the
results are, **###*@@**, less than satisfactory.
    Any rapid fixes or is it back to the trusted hand processing and
work out the TP1020 processing by trial and error. Ian.


Dr. Ian Montgomery,
Histotechnology,
IBLS Support Services,
Thomson Building,
University of Glasgow,
Tel:01413398855
Extn: 8511.


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