1. ABX welcomes the SIMeL April 29th, 2004 ABX Pentra DX 120 Conclusion
Mean features
Leukopoïesis
Erythropoïesis
Thrombopoïesis
Expert validation station
ABX SPS evolution
Conclusion
ABX welcomes
the SIMeL
April 29th, 2004

2. External view Rack loader Reagent compartment Processed tubes
Technical
software
Stat sample Bay
Validation station
Optional slide maker and stainer

3. Main features
45 parameters
Up to 120 samples / hour
Optional and integrated slide maker and stainer
Expert Validation Station

4. Main features 8 types of analyses (with random access) CBC DIFF RET
CBR
DIR
ERB
CBE
SPS(*)
(*): With SPS module

5. Measurement principles overview
IMPEDANCE METHOD:
RBC, PLT, WBC, BASO (specific lyse)
PHOTOMETRY:
HGB - Cyanmethemoglobin or cyanide free
CYTOCHEMISTRY / IMPEDANCE:
5 DIFF + Immature lines
FLUORESCENCE / IMPEDANCE:
NRBC & RET

6. Measurement principles overview
“D.H.S.S.” (HORIBA ABX patented)
Double Hydrodynamic Sequential System
2 measurements points +
Timing control (co%)
Double hydrodynamic focusing to ensure a good measurement
Sequential measurement to ensure that there is no clogging:
-> Correlation Alarm

7. Leukopoïesis exploration

8. Leukopoïesis - WBC count
WBC channel
Baso channel
DIFF channel
Triangulation
Accurate alarms in case of discrepancy between channels

9. Leukopoïesis - WBC count
WBC channel
Double DIFF
channel
Baso channel
ERB channel
QuaDimension
Corrected WBC count if Nrbc presence

10. Focused Flow Impedance
Leukopoïesis - DIFF
Focused Flow Impedance
Measurement of the true cellular volume by impedancemetry after incubation with stabilizing reagent.

11. + Cytochemistry reaction Chlorazol Black Leukopoïesis - DIFF
Lyses of RBC
Stabilises the WBC's in their native forms
Differentially stains the leukocytes.
Staining intensity is proportional to the nucleic acid quantity, to the maturity of this nucleic acid, to the lipids and enzyme quantity.
The absorbance measurement is then proportional to the stain intensity.

12. Tungsten lamp covers almost all wavelengths
Leukopoïesis - DIFF
Light absorption
Exploration of the cell internal structure which is proportional to the stain intensity
Scattering
Tungsten lamp covers almost all wavelengths
Absorption
Scattering
High definition screening
(Cell’s complexity is explored)

13. Leukopoïesis - DIFF
Double Diff Matrix
Absorbance
Resistivity

14. Leukopoïesis - DIFF
Eosinophils
Neutrophils
Lymphocytes
Monocytes

15. Leukopoïesis - DIFF
Atypical Lymphocytes

16. Double Diff Matrix Leukopoïesis - DIFF Large Immature Cells
IMmature Granulocytic line
IMmature Monocytic line
IMmature Lymphocytic line

17. % and # values  High precision of the Diagnosis
Leukopoïesis - DIFF
Abnormal Neutrophil
IMG immature granular
Metamyelocyte
Myelocyte
Promyelocyte
% and # values  High precision of the Diagnosis
Myeloblast

18. % and # values  High precision of the Diagnosis
Leukopoïesis - DIFF
IMM immature monocyte
Promonocyte
Monoblast
% and # values  High precision of the Diagnosis

19. IML immature lymphocyte
Leukopoïesis - DIFF
IML immature lymphocyte
Lymphoblast
Sezary cell
Prolymphocyte
% and # values  High precision of the Diagnosis

20. Additional information provided by Matrix
Leukopoïesis - DIFF
Additional information provided by Matrix
Detection of platelets aggregates
Detection of Nrbc population
Detection of lyse resistant cells
Detection of band cells

21. Evaluations references
ABX PENTRA DX 120: EVALUATION OF THE NEW PARAMETERS
A side-by-side comparison with manual slide review
(Double Matrix / Erythroblasts) and immunophenotyping (Erythroblasts)
Dr. Jose Maria JOU
Servei d’Hemostàsia i Hemoteràpia
Hospital Clínico Universitario de Barcelona, C/ Villarroel nº170, Barcelona, SPAIN.
Myeloid Immaturity and Instrumental Findings
AM. Cenci°, B. Casolari°, M. Maconi*
°Laboratory of Clinical Pathology, AUSL Agency S. Agostino Hospital, Modena; *Laboratory of Chemical Analysis Clinics, Santa Maria Nuova General Hospital, Reggio Emilia.

22. Evaluations references
Evaluation of the ABX Pentra DX 120
Dr. Francis Lacombe
Laboratoire d’Hématologie,
Hôpital Haut-Lévêque, France
New technologies and study of blood cells: the performance of HORIBA ABX PENTRA DX 120
B. Casolaria, M. Maconib, A.M. Cencia
aLaboratory of Clinical Pathology, AUSL Agency S.Agostino Hospital, Modena; bLaboratory of Clinical Pathology, O.I.R.M. Hospital Agency - S.Anna, Turin.
Detection of atypical lymphocyte populations and lymphoid pathology diagnosis in adults and children (ALY alarm)
P. Lemaire, Laboratoire de Longpont-Sur-Orge (91310)
Georges Pompidou European Hospital (75015 Paris), France

23. Erythropoïesis exploration

24. Erythropoïesis - Maturation
The red blood cells line maturation
Proerythroblast
Basophilic Erythroblast
Polychromatophilic Erythroblast
Acidophilic Erythroblast
Reticulocyte
Erythrocyte

25. Measurement principles
NRBC’s count (ERB) 
1. Specific lyse and fluorochrome
a) Lysing of RBC
b) Staining of WBC and NRBC’s with Thiazole orange, a fluorochrome specific
to nucleic acids (reference method).
2. Double Hydrodynamic Sequential System (DHSS)
Flow-cytometer using hydrodynamic focusing before and after the aperture
3. Impedance method
Measurement of the cell volume (“X” axis) 
Argon ion Laser 
4. Fluorescence (and forward scatter)
DNA content (“Y” axis) 

26. Erythropoïesis - Erythroblasts
Fluorescence
WBC
ERB
Volume
% and # values  WBC corrected

27. Evaluations references
ABX PENTRA DX 120 : EVALUATION OF THE NEW PARAMETERS
A side-by-side comparison with manual slide review
(Double Matrix / Erythroblasts) and immunophenotyping (Erythroblasts)
Dr. Jose Maria JOU
Servei d’Hemostàsia i Hemoteràpia
Hospital Clínico Universitario de Barcelona, C/ Villarroel nº170, Barcelona, SPAIN.
Automated Nucleated RBC Counting: Comparison to Flow Cytometry.
Bruce H. Davis*, Kathleen T. Davis*, Esther Tournier+, Karen Becker*
*Trillium Diagnostics, LLC and Maine Medical Center Research Institute, Scarborough, Maine USA
+HoribaABX Diagnostics, Montpellier, France

28. Erythropoïesis - Maturation
The red blood cells line maturation
Proerythroblast
Basophilic Erythroblast
Polychromatophilic Erythroblast
Acidophilic Erythroblast
Reticulocyte
Erythrocyte

29. Erythropoïesis - Reticulocytes
Reticulocyte count
1. Fluorochrome
Reticulocytes are stained according to the RNA they contain.
2. Double Hydrodynamic Sequential System (DHSS)
Flow-cytometer using double hydrodynamic focusing
3. Impedance method
Measurement of the cell volume (“X” axis)
Argon ion Laser
4. Fluorescence (and forward scatter)
RNA content (“Y” axis)

30. Erythropoïesis - Reticulocytes
Volume
Fluorescence
RET
RBC
RET H (RNA +++)
3 maturation stages
function to
the RNA content
RET M (RNA ++)
RET L (RNA +)
IRF -> Immature Reticulocyte Fraction
MFI -> Mean Fluorescence Index
MRV -> Mean Reticulocyte Volume
Monitoring the erythropoietic response
CRC -> Corrected Reticulocyte Count
Classifying anaemias (central/peripheral - generative or non-generative)

31. Evaluations references
The Mean Reticulocyte Volume (MRV) on the HORIBA ABX Pentra 120/DX
as an alternative measurement to the Reticulocyte Haemoglobin Content (CHr™ )
A.H. Roderick – Department of Haematology, University Hospital of Wales, Cardiff. UK
J.M. Jou – Servei d’Hemoterapia, Hospital Clinic, Barcelona. Spain
W.G.B. Rees, E. Thomas – Haematology Department, West Wales General Hospital, Carmarthen. UK
Early diagnosis of iron deficiency anaemia by Mean Reticulocyte Volume (MRV) screening
Dr.Ruud Muusze, clinical chemist, Ziekenhuis Zeeuws Viaanderen, Terneuzen, Holland.

32. Erythropoïesis - Maturation
The red blood cells line maturation
Proerythroblast
Basophilic Erythroblast
Polychromatophilic Erythroblast
Acidophilic Erythroblast
Reticulocyte
Erythrocyte

33. Erythropoïesis - Red Blood Cells
Specific quantitative flags:
Anemia: Hgb
Polyglobulia: RBC
Cold agglutinin: MCHC
Micro/macrocytosis: MCV
Anisocytosis: RDW
Reference methods of measurement  High precision of the Anemias and RBC pathologies

34. Erythropoïesis - Red Blood Cells
Specific morphology flags:
MIC: presence of microcytic population
MAC: presence of macrocytic population
No (noise): presence of lyse resistant cells
RBC morphology  confirmation of the Diagnosis
 detection of abnormal RBC pop.

35. Erythropoïesis - Red Blood Cells
Specific and precise alarm MIC (3 levels)
microcytes, poïkilocytes, schistocytes, spherocytes, drepanocytes, Joly bodies …
Specific and precise alarm MAC (3 levels)
macrocytes, poïkilocytes, ponctuate basophilia...

36. Thrombopoïesis exploration

37. Normal platelets population
Thrombopoïesis
Normal platelets population
Specific MIC flag

38. Thrombopoïesis
Platelet aggregates
Optical detection

39. Thrombopoïesis
Macroplatelets
Macroplatelet detection

40. Evaluation references
Evaluation of Platelets count below /L: Hematologic Analyzers
ABX Pentra DX 120 versus platelet reference method
Dr. Jose Maria JOU
Servei d’Hemostàsia i Hemoteràpia
Hospital Clínico Universitario de Barcelona, C/ Villarroel nº170, Barcelona, SPAIN.

41. Evaluation references
Am J Clin Pathol 115(3): , © 2001 American Society of Clinical Pathologists, Inc

42. Evaluation references

43. Expert validation station

44. Specs & functionality
Dedicated PC
Windows XP, CD-ROM burner
Onboard user manual
Mono or bi-directional host connection
Connection of 3 ABX analyzers
Query mode
Download capability

45. _ Automatic validation _ Messages, comments….
Specs & functionality
User defined rules
_ Rerun capability
_ Reflex testing
_ Automatic validation
_ Messages, comments….
Bio-technical validation with :
Patient history, delta check
Manual entry
Recalculation capability
Old
New D+ D-
Delta check

46. Specs & functionality
results
1 matrix
3 histograms
Zoom capability

47. Specs & functionality
Hematovision IV included
A cell cytology library

48. SPS EVOLUTION

49. Throughput : 120 stained slides per hour Sample volume: 50µl
Specification
Throughput : 120 stained slides per hour
Sample volume: 50µl
Stain : MGG, Wright, Wright-Giemsa
User defined smear criteria
Smearing profile function to each individual sample and to each hematology parameter
Automatic maintenance

50. When dispensed, slides are twisted to avoid blockages
Technology
Slide stacker capacity of 160 slides
When dispensed, slides are twisted to avoid blockages
Rack stacker capacity of 8 racks
Located outside, slides and racks are easily reloaded at any time

51. Titanium coated wedge :
Technology
Titanium coated wedge :
No change, no adjustment
Smearing tape :
Moves forward between each sample to avoid contamination
and to avoid a complex mechanism to clean and dry the wedge

52. User’s defined smearing profile : Technology
- Blood drop volume adjustable by increment of 0,1 µl
- 2 smearing angles (40° or 45°)
- Waiting time adjustable by increment of 1ms
- Smearing speed adjustable by increment of 1step/ms

53. Technology

54. Slide identification :
Technology
Slide identification :
3 lines of 14 alpha-numerical characters
Drying process :

55. Individual staining wells :
Technology
Individual staining wells :
Use of fresh stains ensures a good staining stability
User defined stain volume
Automatic stain dilution
Staining steps adjustable by increment of 30s
Maximum staining duration of 16mn

56. 20 slides can collected in one rack
Technology
Manual mode :
Pre-smeared slide:
Bone marrow
Biologic fluid
Cyto-centrifuging
Leuko-concentration
Slide outlet :
20 slides can collected in one rack

57. Manual smear SPS Evolution Technology Manual identification
Irregular blood films
Irregular staining
Automatic identification
Homogenous blood films
Standardization of the staining

58. ABX Pentra DX 120
Conclusion
Dr. Echevarne laboratory

59. Laboratorios Dr. F. Echevarne
Rack
Pentra DX+SPS
Pentra DX
Consolidate Storage Data Base
SERVER
Validation Station
Medical Validation Station
ASTM
ASTM “Host Query”
HORIBA ABX SAT5000
Automatic Tube Sorter & Storage
Host
Computer
Pentra DX Cell
Central Laboratory
>4.500 Samples/day
Satelit Labs. & Clinics
1 site
Pentra 80
7 Clinics
2 Satelite Labs.
6 Satelite Labs.
1 Clinics
Pentra 60 C+
5 Clinics
18 x Pentra 80
16 x Pentra 60 C+

60. HORIBA ABX ambitious goals will be reached by becoming the
Conclusion
HORIBA ABX is differentiating itself from the other “suppliers of systems” in being a “partner providing solutions” for the laboratory.
HORIBA ABX ambitious goals will be reached by becoming the
“preferred partner” of its customers worldwide.

61. Thank you
ABX welcomes
the SIMeL
April 29th, 2004